Highly potent inhibitors of TNF-α production. Part I: Discovery of new chemical leads and their structure-activity relationships

Bioorganic & Medicinal Chemistry10 (2002) 3757–3786

Highly Potent Inhibitors of TNF-ꢀ Production. Part I:

Discovery of New Chemical Leads and Their

Structure–Activity Relationships

Toshiaki Matsui,^a,* Takashi Kondo,^bYoshitaka Nishita,^aSatoshi Itadani,^b

Shingo Nakatani,^bNagashige Omawari,^cMasaru Sakai,^bShuichi Nakazawa,^c

Akihito Ogata,^bHideaki Mori,^aKouichiro Terai,^bWataru Kamoshima,^b

Hiroyuki Ohno,^bTakaaki Obata,^bHisao Nakai^band Masaaki Toda^b

^aFukui Research Institute, Ono Pharmaceutical Co., Ltd., Technoport, Yamagishi, Mikuni, Sakai, Fukui 913-8638, Japan

^bMinase Research Institute, Ono Pharmaceutical Co., Ltd., Shimamoto, Mishima, Osaka 618-8585, Japan

^cHeadquarters, Ono Pharmaceutical Co., Ltd., Doshoumachi, Chuou, Osaka 541-8526, Japan

Received 26 June 2002; accepted 5 August 2002

Abstract—Discoveryof new chemical leads of inhibitors for TNF- a production starting from the chemical modiﬁcation of 1 is

reported. Further biological studies of 1 to disclose the site of its action stronglysuggested that 1 inhibits LPS-induced TNF-a

expression in the liver and spleen of mice. Structure–activityrelationships (SARs) are also discussed and full details including the

chemistryare reported.

Introduction

As one of the keycytokines in diseases such as rheumatoid

arthritis (RA), multiple sclerosis, cachexia, sepsis, ulce-

rative colitis, congestive heart failure, inﬂammatorybowel

disease and Crohn’s disease, TNF-a^1ꢀ3is thought to be at

or near the top of hierarchyof all cytokines that playa

role in these diseases.^4ꢀ9This important role has been

Figure 1. New inhibitors of TNF-a production.

demonstrated bystudies illustrating that blockage of

TNF-a activityin snyovial cells from rheumatoid

arthritis (RA) patients inhibits the production of

another important cytokines (IL-1 and other cytokines)

which promote inﬂammation. Thus, TNF-a plays a

central role in the initiation and maintenance of the

diseases as described above. Because of its therapeutic

potential, much attention has been paid to the develop-

ment of a small molecule, which inhibits TNF-a

production.^10ꢀ13In one of our preceding papers,^14,15we

reported on 2-(octanoylamino)-2-phenylethyl disodium

phosphates 1–2 as a new class of chemical lead (Fig. 1).

We report here the full details of the discoveryprocess

for the chemical leads and further biological studies of 1

to disclose its site of action. Structure–activityrelation-

ships (SARs) are also discussed.

Chemistry

Synthesis of all the compounds listed in the Tables 1–7

are described in Schemes 1–11. As outlined in Scheme 1,

N-[(1R)-2-hydroxy-1-phenylethyl]acylamides 60a–r,t and

N-[(1R)-2-hydroxy-1-phenylethyl]octane-1-sulfonamide

60s prepared from their corresponding aminoalcohols,

were converted to dibenzyl phosphates 61a–t, deprotec-

tion of which provided 62a–t. Disodium phosphates 1,

3–20 and 22 were prepared from their corresponding

*Corresponding author. Tel.: +81-756-82-6161; fax: +81-756-82-

8420; e-mail: to.matsui@ono.co.jp

PII: S0968-0896(02)00381-4

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T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

Table 2. Biological evaluation of hetero atom-containing N-acyl

derivatives 14–22

Table 1. Optimization of the N-acyl moiety

Compd

R

Inhibition of TNF-a production

ID₅₀(mg/kg, iv)

Compd

R

Inhibition of TNF-a production:

ID₅₀(mg/kg, iv)

Mice

1.1

Rats

3.2

Mice

Rats

3

4

5

6

1

7

8

Me

n-Pr

(28)^a

(17)^a

30

(21)^b

(37)^b

50

4.5

3.0

N.T.^c

14

n-C₅H₁₁

n-C₆H₁₃

n-C₇H₁₅

n-C₁₀H₂₁

–Ph

2.8

0.8

15

16

17

18

19

20

21

22

(14)^a

(12)^a

(1)^a

N.T.^c

(38)^b

10.5

(50)^a

(32)^a

9

(36)^b

N.T.^c

10

11

(27)^a

(56)^a

N.T.^c

(34)^a

(58)^a

(ꢀ10)^a

N.T.^c

12

(59)^a

4.2

N.T.^c

8.0

13a (less polar)

13b (more polar)

2.4

17

^aInhibition % at 10 mg/kg, iv.

^bInhibition % at 30 mg/kg, iv.

^cN.T.: not tested.

^aInhibition % at 30 mg/kg, iv.

^bInhibition % at 10 mg/kg, iv.

^cN.T.: Not tested.

free acid forms, respectivelybytreatment with NaOH

aq in EtOH. The (2R)-pentylsulfonylethanoyl derivative

21 was prepared from its sulﬁde derivative 62t byan

oxidation reaction with OXONE¹followed bytreat-

ment with NaOH aq in EtOH. As described in Scheme

2, 23a–26c and 47 were synthesized from their corre-

sponding methyl aminophenylacetate 63a–k.¹⁶N-Acy-

lation of 63a–k with an octanoyl chloride followed by

the reduction with LiBH₄in THF provided 64a–k.

Protected phosphates 65a–k were prepared from their

corresponding N-acylaminoalchols 64a–k, respectively.

Deprotection of 65a–k aﬀorded the free acid forms 66a–k,

which were converted to their disodium phosphates

23a–26c and 47, respectively. As outlined in Scheme 3,

compounds 53–57 were prepared from 67a–e according

to essentiallythe same procedures as described in

Scheme 2. As described in Scheme 4, compounds 27–40

were synthesized from a common starting material 70.

O-Alkylation of the phenolic alcohol of 70 followed by

the reduction with LiBH₄in THF aﬀorded inter-

mediates 71a–m, which were converted to 27–38 and 40

according to the same procedures as described before.

Catalytic hydrogenation of the O-benzyl moiety of 71m

followed by O-alkylation with ethyl bromoacetate

aﬀorded 72, which was converted to 39 bythe following

Table 3. Biological evaluation of compounds 23a–26 possessing a

substituted phenyl moiety

Compd

X

Inhibition of TNF-a production

in rats ID₅₀(mg/kg. iv)

23a

23b

23c

24a

24b

24c

25a

25b

25c

26

2-OMe

3-OMe

4-OMe

2-Me

3-Me

4-Me

2-Cl

3-Cl

4-Cl

3.5-OMe

(55)^a

0.5

(34)^a

2.0

5.3

(44)^a

(46)^a

5.2

(ꢀ118)^a

3.8

^aInhibition % at 10 mg/kg, iv.

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3759

successive procedures: (1) treatment with i-Pr₂NP(OB-

17a,b

Table 6. Biological evaluation of the opticallyactive forms 1–2 and

51–52

n)₂followed byan oxidation reaction with

m-CPBA; and (2) deprotection byhydrogenation fol-

lowed bytreatment with NaHCO aq. As described in

3

Scheme 5, compounds 41 and 42 were prepared from

71m. Compound 71m was converted to 73 bythe fol-

lowing successive procedures: (1) protection of the

hydroxy group; (2) hydrogenolysis of the benzyl ether

moiety; (3) triﬂuoromethane sulfonylation; and (4) pal-

ladium-catalyzed carbonylation by the conventional

Compd

R₁

R₂

X

H

Inhibition of

TNF-a

production ID₅₀

(mg/kg, iv) rats

Table 4. Biological evaluation of meta-alkoxyphenly derivatives

27–40

1

H

3.0

2

OMe

H

0.26

10.0

1.6

51

52

H

Compd

R

Inhibition of TNF-a

production in rats

ID₅₀(mg/kg, iv)

OMe

27

28

29

30

31

32

H

Et

ⁿPr

iPr

ⁿBu

ⁱBu

1.4

0.2

0.7

0.1

(25)^a

3.3

Table 7. Biological evaluation of the miscellaneous aromatic deriva-

tives 53–59

33

(24)^a

34

35

ⁿPentyl

ⁿHex

(23)^a

(ꢀ6)^a

0.9

36

37

(7)^a

Compd

AR

Inhibition of TNF-a

production in rats

ID₅₀(mg/kg, iv)

38

39

40

CH₂CON(Me)₂

CH₂COOEt

CH₂OCH₃

(36)^a

(19)^a

1.5

^aInhibition % at 10 mg/kg, iv.

53

3.9

Table 5. Biological evaluation of compounds 41–50 possessing a

miscellaneous meta-substituent

54

55

(6.5)^a

3.8

Compd

X

Inhibition of TNF-a

production in rats

ID₅₀(mg/kg, iv)

56

57

58

59

(16)^a

(ꢀ5)^a

(17)^a

(56)^a

41

42

43

44

45

46

47

48

49

50

CH₂OH

CH₂OCH₃

COONa

COOCH₃

CH₂COONa

CH₂COOCH₃

SMe

SⁱPr

SO₂CH₃

N(Na)SO₂CH₃

3.0

4.1

(37)^a

0.4

(26)^a

4.2

0.3

0.2

(31)^a

(36)^a

^aInhibition % at 10 mg/kg, iv.

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T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

Scheme 1. Synthesis of compound 1 and 3–22. Reagents: Method A: (a) (i) n-BuLi, THF; (ii) (BnO)₂P(O)Cl; (b) H₂, Pd–C, MeOH; (c)

NaOHaq, EtOH; Method B: (a) (i) LDA, THF; (ii) [(BnO)₂P(O)]₂O; (b) H₂, Pd–C, MeOH; (c) NaOHaq, EtOH; Method C: (a) (i) NaH,

THF; (ii) (^tBuO) ₂P(O)Br; (b) TFA; (c) NaOHaq, EtOH; Preparation of 21: (d) (i) TFA; (ii) OXONE¹; (iii) NaOHaq, EtOH.

Scheme 2. Synthesis of compounds 23a–26 and 47. Reagents: (a) n-C₇H₁₅COCl, Py, or Et₃N, CH₂Cl₂, or n-C₇H₁₅COCl, satd NaHCO₃, dioxane; (b)

LiBH₄, THF; Method A (c–g): (c) (i) n-BuLi, THF, (ii) (BnO)₂P(O)Cl; (d) H₂, Pd–C, MeOH; (e) NaOHaq, EtOH; Method C (f, g, e): (f) (i) NaH,

benzene-THF, (ii) (^tBuO)₂P(O)Br; (g) TFA; (h) OXONE¹, MeOH.

method.¹⁸Lithium borohydride reduction of the ester

moietyof 73 followed bythe protection of the formed

hydroxymethyl moiety with chloromethyl methyl ether

(MOMCl) and desilylation with tetrabutylammonium

ﬂuoride (TBAF) provided 74. Lithium borohydride

reduction of the ester moietyof 73 followed by O-methy-

lation and deprotection aﬀorded 75. Phosphates 76 and 77

were prepared from 74 and 75, respectivelybythe follow-

ing successive procedures: (1) phosphinylation;¹⁹(2) oxi-

dation with m-CPBA; (3) deprotection with 50% Me₂NH

in EtOH; and (4) acidiﬁcation. Compounds 76 and 77

were converted to 41 and 42, respectivelyaccording to

the same procedures as described before. Synthesis of

43–46 is described in Scheme 6. Deprotection of the

TBDMS group in 73 gave 78, which was phosphory-

lated to 43 and 44 bythe following successive proce-

dures: (1) phosphinylation; (2) oxidation with m-CPBA;

(3) deprotection with DBU; and (4) treatment with

NaOH aq or NaHCO₃aq. O-Benzylation of 78 fol-

lowed bythe alkaline hydrolysis aﬀorded 79, which was

converted to 80 bythe Arndt-Eistert C1 homologation

reaction.²⁰Hydrogenolysis of the benzyl ether of 80

followed byessentiallythe same procedures as described

above provided 45 and 46. Compound 48 was prepared

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3761

Scheme 3. Synthesis of compounds 53–57. Reagents: (a) n-C₇H₁₅COCl, Py, CH₂Cl₂; (b) LiBH₄, THF; Method A (c–e): (c) (i) n-BuLi, THF,

(ii) (BnO)₂P(O)Cl; (d) H₂, Pd–C, MeOH; (e) NaOHaq, EtOH.

Scheme 4. Synthesis of compounds 27–40. Reagents: (a) RX, K₂CO₃, DMF or acetone; (b) LiBH₄, THF; Method A (c–e): (c) (i) n-BuLi, THF,

(ii) (BnO)₂P(O)Cl; (d) H₂, Pd-C, MeOH; (e) NaOHaq, EtOH; (f) BrCH₂COOEt, KI, K₂CO₃, DMF; Method D (g, h, d, i): (g) i-Pr₂NP(OBn)₂,

tetrazole, CH₃CN; (h) m-CPBA, CH₂Cl₂; (i) NaHCO₃aq, EtOH; (j) 6M HCl, MeOH; Preparation of 71j and 71g: (k) ROH, DEAD, Ph₃P, THF.

Scheme 5. Synthesis of compounds 41 and 42. Reagents: (a) TBDMSCl, imidazole, DMF; (b) H₂, Pd–C, MeOH; (c) Tf₂O, Py; (d) Pd(OAc)₂, DPPP,

CO, Et₃N, MeOH, DMSO; (e) LiBH₄, THF; (f) MOMCl, i-Pr₂NEt, CH₂Cl₂; (g) NaH, MeI, THF; (h) TBAF, THF; Method E (i–m):

(i) i-Pr₂NP(OCH₂CH₂CN)₂, tetrazole, CH₃CN; (j) m-CPBA, CH₂Cl₂; (k) 50% Me₂NH, EtOH; (l) c-HCl; (m) NaOHaq, EtOH.

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T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

Scheme 6. Synthesis of compounds 43–46. Reagents: (a) TBAF, THF; Method C (b–e or b–d, f): (b) i-Pr₂NP(OCH₂CH₂CN)₂, tetrazole, CH₃CN;

(c) m-CPBA, CH₂Cl₂; (d) DBU, THF; (e) NaOHaq; (f) satd NaHCO₃, EtOH; (g) Ag₂O, BnBr, DMF; (h) 2N-NaOH, THF, EtOH; (i) (i) (COCl)₂,

DMF, toluene; (ii) CH₂N₂, Et₂O; (iii) AgOAc, Et₃N, MeOH; (j) H₂, Pd–C, MeOH, Method D (k–l, e or k–l, f): (k) i-Pr₂NP(OBn)₂, tetrazole,

CH₃CN; (l) m-CPBA, CH₂Cl₂.

Scheme 7. Synthesis of compound 48. Reagents: (a) NaH, ClC(S)NMe₂, DMF; (b) 235 ^ꢁC, Ph₂O; (c) LiBH₄, THF; (d) KOH, MeOH; (e) PrI,

i

K₂CO₃, DMF; Method C (f–h): (f) NaH, (^tBuO)₂P(O)Br, THF; (g) TFA, CH₂Cl₂; (h) NaOHaq, EtOH.

Scheme 8. Synthesis of compound 50. Reagents: (a) TBDMSCl, imidazole, DMF; (b) H₂, Pd–C, MeOH; (c) Tf₂O, Py; (d) Ph₂CNH, Pd(OAc)₂,

.

BINAP, Cs₂CO₃, toluene; (e) HONH₂HCl, AcONa, MeOH; (f) MsCl, Py; (g) TBAF, THF; Method D (h–j): (h) i-Pr₂NP(OBn)₂, tetrazole, CH₃CN;

(i) m-CPBA, CH₂Cl₂; (j) NaOHaq, EtOH.

from 70 as outlined in Scheme 7. Compound 70 was

converted to 81 bythe following successive procedures:

(1) replacement of the phenol group with a thiol group

method.²²Compound 83 was converted to 84 bythe fol-

lowing successive procedures: (1) selective hydrolysis of

the imino moietyfollowed by N-methanesulfonylation;

and (2) desilylation with TBAF. Conversion of 84 to 50

was accomplished bythe above-mentioned procedures.

As outlined in Scheme 9, compounds 2 and 53 were

21

bythe conventional method;

(2) lithium borohydride

reduction of the ester group; and (3) alkaline hydrolysis

followed by S-alkylation. Compound 81 was converted

to 48 bythe same procedures as described before.

Compound 50 was prepared from 71m as described in

Scheme 8. Compound 71m was transformed to 82

according to the same procedures as described in Scheme

5. Compound 82 was converted to 83 bythe reported

23a,b

prepared from the opticallyactive aminoalcohols,

85 and 87, respectivelyaccording to the same procedure as

described before. As described in Scheme 10, compound

52 was prepared from 89, which was transformed to a

24

phosphate 90 bythe conventional method. Compound

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3763

Scheme 9. Synthesis of compounds 2 and 53. Reagents; (a) n-C₇H₁₅COCl, satd NaHCO₃aq, THF; Method A (b–d): (b) (i) n-BuLi, THF; (ii)

(BnO) ₂P(O)Cl; (c) H₂, Pd–C, MeOH; (d) NaOHaq, EtOH.

Scheme 10. Synthesis of compound 52. Reagents: Method F (a–e): (a) (Cl₃CCH₂O)₂P(O)Cl, Py; (b) TFA, CH₂Cl₂; (c) n-C₇H₁₅COCl, Py, CH₂Cl₂;

(d) Zn, Py, AcOH; (e) NaOHaq, EtOH.

Scheme 11. Synthesis of compounds 58 and 59. Reagents: (a) n-C₇H₁₅COCl, NaHCO₃aq, THF; (b) LiBH₄, THF; Method A (c–e): (c) (i) n-BuLi,

THF; (ii) (BnO)₂P(O)Cl; (d) H₂, Pd-C, MeOH; (e) NaOHaq, EtOH.

90 was converted to 52 according to the following suc-

cessive procedures: (1) deprotection of the N-tert-butyl-

oxycarbonyl group; (2) N-acylation with octanoyl

chloride; (3) reductive deprotection of the trichloroethyl

group; and (4) treatment under alkaline conditions. As

outlined in Scheme 11, heteroaromatic derivatives 58 and

59 were prepared from 91a²⁵and 91b,²⁶respectively.

N-Acylation followed by reduction aﬀorded 92a and

92b, which were converted to 58 and 59 bythe usual

methods, respectively.

using a commerciallyavailable kit. Test compounds

were intravenously(iv) administered prior to the iv

injection of LPS. Potencyof the test compounds was

evaluated bytheir ID

values (the dosage required to

50

inhibit plasma TNF-a production by50%) or percen-

tage of inhibition at the maximum dosages administered

(10 mg/kg or 30 mg/kg, iv).

As described below, optimization of the N-acyl moiety

(Tables 1 and 2), the substituent on the aromatic moiety

(Tables 3 and 5) and biological evaluation of the opti-

callyactive forms (Table 6) were studied. An attempt to

ﬁnd another aromatic moietywhich could be sub-

stituted for the phenyl moiety of 1 was also carried out,

the result of which is described in Table 7.

Results and Discussion

A series of 2-(acylamino)-2-phenylethyl disodium phos-

phates was synthesized and biologically evaluated for

their abilityto inhibit LPS-induced plasma TNF- a pro-

duction in mice and rats. Plasma TNF-a production

was determined 90 min after LPS injection byELISA

In the course of our screening program for the TNF-a

inhibitors, a new class of compounds, 2-(acylamino)-2-

phenylethyl disodium phosphate derivatives 1 and 2

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T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

were discovered to be highlypotent inhibitors of TNF- a

production. Our chemical modiﬁcation was started with

the optimization of the N-acyl chain in 1 followed by

the introduction of an alkoxygroup into the meta-posi-

tion of the phenyl moiety. Due to the eﬀectiveness of the

screening process, the biological evaluations were car-

ried out in mice in the earlier stages of the project and in

rats in the later stages.

evaluations with mice than with rats.

Optimization of the substituents on the phenyl moiety

was carried out using racemic compounds for their syn-

thetic reasons as illustrated in Tables 3–5. As described

in Table 3, introduction of a methoxygroup into the

phenyl moiety of (ꢂ)-1 aﬀorded 23a–c. Of these, the

meta-isomer 23b exhibited the most potent activityin

the evaluation with rats. The inhibitoryactivityof the

ortho-isomer 23a was more potent than that of para-

isomer 23c. Introduction of a methyl group into the

phenyl moiety of (ꢂ)-1 instead of a methoxygroup

aﬀorded 24a–c. Among them, the ortho-isomer 24a

exceptionallydemonstrated the most potent activit.y

The meta-isomer 24b showed more potent inhibitory

activitythan para-isomer 24c but less potent activity

than 24a. Introduction of a chloro group into the

phenyl moiety in (ꢂ)-1 provided 25a–c. Of these, the

meta-isomer 25b again demonstrated the most potent

activity. The ortho-isomer 25a also showed more potent

activitythan the para-isomer 25c. Thus, meta-isomer

tended to show the most potent activityamong the each

of the classes. The ortho-isomers 23a, 24a and 25a

always showed more potent activity than their corre-

sponding para-isomers 23c, 24c and 25c, respectively.

Introduction of another meta-methoxygroup into the

phenyl moiety of 23b aﬀorded 26 with a nearlyeight

times less potent ID₅₀value.

As described in Table 1, good ID₅₀values were

obtained with 6, 1 and 13a–b. The importance of the

role played by the N-acyl moiety in the potency of

these compounds was clariﬁed bythe marked reduction

in the inhibitoryactivityof 3–5 and 7–10. As illustrated

in 1 and 3–7, the length of the N-acyl side chain was

optimized at the N-octanoyl moiety. Compounds 8–10

possessing benzoyl, branched alkanoyl and 5-phenyl-

pentanoyl moieties, respectively aﬀorded a marked

reduction in their inhibitoryactivitycompared with 1.

To block a predicted inactivation by rapid hydrolysis

of the N-acyl chain, a 2,2-dimethyl group and a 2,2-

trimethylene group were introduced into the optimized

N-octanoyl moiety of 1 aﬀorded 11 and 12 with a

reduced activity. Introduction of a methyl group into

the same position aﬀorded two diastereomers 13a (less

polar isomer) and 13b (more polar isomer). The inhi-

bitoryactivityof the more polar isomer 13b was more

potent than that of the less polar isomer 13a. Thus, the

SARs obtained in mice were roughlyretained with the

evaluation in rats.

As described in Table 4, further optimization of the

meta-alkoxymoietywas carried out. The meta-hydroxy

derivative 27 demonstrated an ID₅₀value of 1.4 mg/kg,

iv. The ID₅₀values of the ethoxyderivative 28 and iso-

propyloxy derivative 30 were the most potent among

the test compounds 27–40. Although the ID₅₀values for

29 and 36 showed that theywere still potent. The iso-

butyl derivative 32 demonstrated moderate potency,

however compounds 31, 33–35 and 37 showed a marked

decrease in inhibitoryactivitypresumablybecause of the

more bulky meta-alkoxygroups. Heteroatom-containing

meta-alkoxyderivatives 38–40 were also synthesized and

biologicallyevaluated. Of these, the methoxymethyl deri-

vative 40 exhibited the most potent ID₅₀value.

Introduction of a heteroatom into the N-acyl chain

provided compounds 14–22 as illustrated in Table 2.

The n-hexyloxycarbonyl derivative 14 showed nearly

same potencyas 1 on its evaluation in both mice and

rats. Other oxygen-containing N-acyl derivatives 15–18

showed less than 50% inhibition at a dose of 30 mg/kg,

iv (mice). The urea derivative 19 demonstrated less

potent activitycompared with the corresponding ure-

thane derivative 14. N-Sulfonyl derivative 20 did not

show anyactivityat 30 mg/kg, iv (mice). The 2-pentyl-

sulfonylmethyl derivative 21 showed less than 50%

inhibition at 10 mg/kg, iv (rats) while the 2-pen-

tylthioethanoyl derivative 22 had an ID₅₀value of 10.5

mg/kg (rats). Thus, the N-carbonyl moiety is one of the

structural requirements for potent inhibitoryactivityas

illustrated in 1 and 14.

As described in Table 5, other meta-substituted deriva-

tives 41–50, the meta-position of which was substituted

with carbon, sulfur and nitrogen atoms, were also syn-

thesized and biologicallyevaluated. Of these, a methyl

ester, as well as methylthio and isopropylthio deriva-

Replacement of one of the N-acyl carbons with an oxy-

gen atom provided 14–18. The increased hydrophilicity

in the N-acyl side chains of 15–18 was thought to be

deleterious for their potent inhibitoryactivit.y The

N-hexyloxycarbonyl moiety of 14 was thought to be an

isoster of the N-octanoyl moiety of 1. The lower activ-

ityof the urea derivative 19 was thought to be due to

the more polar propertyof its urea moietycompared

with that of the urethane moietyof 14. Replacement of

the carbon atom at position-3 with sulfonyl or sulﬁde

was also found to be deleterious for the inhibitory

activity.

tives 44, 47 and 48 demonstrated verypotent ID

50

values. Hydroxymethyl, methoxymethyl and methoxy-

carbonylmethyl derivatives 41, 42 and 46 demonstrated

moderate ID₅₀values (3–4 mg/kg, iv). The remaining

compounds 43, 45, 49 and 50 showed less than 50%

inhibition at the dose of 10 mg/kg, iv. Carboxyl, car-

boxylmethyl, methyl sulfonyl and methylsulfonyl amino

groups were found to be deleterious for increasing the

inhibitoryactivity.

As described in Table 6, opticallyactive derivatives 1–2

and 51–52 were synthesized and biologically evaluated.

The inhibitoryactivityof the newlydiscovered chemical

lead 1 exhibited inhibitoryactivityfor TNF- a production

According to the data obtained so far, the ID₅₀values

of these compounds were normallymore potent in the

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3765

at 3.0 mg/kg, iv in rats. The meta-methoxyderivative 2

showed a nearlyten times more potent ID value (0.26

50

mg/kg, iv in rats) than 1. Their corresponding enan-

tiomers 51 and 52 demonstrated nearlythree times and

six times less potent ID₅₀values, respectively. Thus the

(R)-conﬁguration of 1 and 2 was thought to be a

structural requirement for their more potent activity

compared with the (S)-conﬁguration of 51 and 52.

A biological evaluation of the miscellaneous aromatic

derivatives 53–59 is outlined in Table 7. Replacement of

the phenyl moiety in (ꢂ)-1 with a 1-naphtyl moiety

aﬀord 53 which retained the potent activitywhile the

potencywas markedlydecreased in the evaluation of the

corresponding 2-naphthyl derivative 54. A similar ten-

dencyto the above-mentioned result was obtained in the

biological evaluation of 55 and 56. As such, 55 demon-

strated nearlythe same potencyas 53 while 56, which

corresponds to 54, demonstrated much less potency

than 55. The 2-thienyl derivative 57 did not exhibit any

inhibitoryactivityat the dose of 10 mg/kg, iv although a

thienyl moiety is very often used as an isoster of a

phenyl moiety in medicinal chemistry.

Figure 2. Evaluation of TNF-a expression in mouse liver byRT-PCR.

M: DNA molecular weight markers. Lane 1: saline; lane 2: LPS (5 mg/

kg, ip); lane 3: LPS (5 mg/kg, ip)+compound 1 30 mg/kg, ip (1 min

before LPS injection); lane 4: LPS (5 mg/kg, ip)+PGE₂1 mg/kg, ip (1

min before LPS injection).

Pyridine derivatives 58–59 tended to show inhibitory

activityalthough their potencywas not so high. Com-

pound 59 showed more than 50% inhibition at 10 mg/kg,

iv while 58 exhibited 17% inhibition at the same dose.

LPS, an inﬂammatoryinducer, stimulates TNF- a

mRNA and protein expression. It has been reported

that signiﬁcant increases of TNF-a mRNA expression

were seen in spleens and livers of LPS-treated animals.^27aꢀc

As described in Table 1, compound 1 showed a potent

inhibitoryeﬀect on increase of plasma TNF- a produc-

tion in LPS-treated mice. To elucidate mechanism of the

inhibitoryeﬀect of 1 on TNF-a protein expression we

investigated an eﬀect of 1 on TNF-a mRNA induction

in spleens and livers of LPS-treated mice. As shown in

Figures 2 and 3, compound 1 (line 3) signiﬁcantlysup-

presses LPS-induced increase of TNF-a production at

the mRNA level as well as PGE₂(line 4), which was

used as the positive control. The mechanism leading to

inhibition on plasma TNF-a production in LPS-treated

mice likelydepends on suppression of TNF- a mRNA

induction.

Figure 3. Evaluation of TNF-a expression in mouse spleen byRT-

PCR. M: DNA molecular weight markers. Lane 1: saline; lane 2: LPS

(5 mg/kg, ip); lane 3: LPS (5 mg/kg, ip)+compound 1 30 mg/kg, ip (1

min before LPS injection); lane 4: LPS (5 mg/kg, ip)+PGE₂1 mg/kg,

ip (1 min before LPS injection).

of 1 stronglysuggested that the inhibitoryactivityof

1

in LPS-induced TNF-a production takes place at the

level of TNF-a expression in the liver and spleen of

mice. The ﬁndings from the present studywill be useful

for further optimization of the newlydiscovered chemi-

cal lead 2. Lead optimization and oral activityof repre-

sentative test compounds will be reported in the

following paper in this journal.

Conclusion

In summary, we have discovered a new series of inhibi-

tors of TNF-a production. A number of the compounds,

2-(acylamino)-2-phenylethyl phosphate derivatives, most

notably 2, 23b, 28, 30, 44, 47 and 48, were excellent

inhibitors. Biological evaluation of the opticallyactive

derivatives clearlyshows that ( R)-enantiomers 1 and 2

are more potent inhibitors than their corresponding

(S)-enantiomers 51 and 52, respectively. According to

the biological evaluation of the miscellaneous aromatic

derivatives 53–59, 53 and 55 exhibited nearlythe same

potencyas 1. Based on this result, the 2-naphtyl and 2,3-

methylenedioxyphenyl moieties could be a surrogate for

the phenyl moiety of 1. Our further biological evaluation

Experimental

General directions

Analytical samples were homogeneous as conﬁrmed by

TLC, and aﬀorded spectroscopic results consistent with

the assigned structures. All ¹H NMR spectra were taken

on a Varian Gemini-200, VXR-200s or Mercury300

spectrometer. MS spectra were obtained on a Hitachi

M1200H, JMS-DX303HF or PerSeptive Voyager

3766

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

Elite spectrometer. Matrix assisted laser desorption

ionization-time of ﬂight high-resolution mass spectra

(MALDI-TOF HRMS) were obtained on a PerSeptive

Voyager Elite spectrometer. IR spectra were measured

on a Perkin–Elmer FT-IR 1760X or Jasco FT/IR-430

spectrometer. Melting points were uncorrected. Optical

rotations were measured on a Jasco DIP-1000 polari-

meter. Column chromatographywas carried out on

silica gel (Merck silica gel 60 (0.063–0.200 mm) or Fuji

Silysia FL60D). Thin layer chromatography was per-

formed on silica gel (Merck TLC plate, silica gel 60

F₂₅₄). HPLC analyses were performed with a LaChrom

(L-7400 UV Detector, L-7100 pump: HITACHI). The

following abbreviations for solvents and reagents are used:

THF, tetrahydrofuran; EtOAc, ethyl acetate; MeOH,

methanol; EtOH, ethanol; DMF, dimethylformamide;

N-[(1R)-2-Hydroxy-1-phenylethyl]hexanamide (60c). Yield,

1

70%; TLC R_f=0.40 (n-hexane/EtOAc, 1/2); H NMR

(300 MHz, CDCl₃) d 7.40–7.25 (m, 5H), 6.23 (brd,

J=6.3 Hz, 1H), 5.05 (m, 1H), 3.86 (brd, J=4.8 Hz, 2H),

2.23 (t, J=7.5 Hz, 2H), 1.67–1.58 (m, 2H), 1.41–1.25

(m, 4H), 0.88 (brt, J=6.9 Hz, 3H).

N-[(1R) -2-Hydroxy-1-phenylethyl]heptanamide (60d).

1

Yield, 99%; TLC R_f=0.46 (n-hexane/EtOAc, 1/2); H

NMR (300 MHz, CDCl₃) d 7.40–7.25 (m, 5H), 6.26

(brd, J=6.6 Hz, 1H), 5.05 (m, 1H), 3.86 (brd, J=5.1

Hz, 2H), 2.23 (t, J=7.5 Hz, 2H), 1.67–1.58 (m, 2H),

1.41–1.25 (m, 6H), 0.87 (brt, J=6.9 Hz, 3H).

N-[(1R)-2-Hydroxy-1-phenylethyl]benzamide (60g). Yield,

1

68%; TLC R_f=0.40 (n-hexane/EtOAc, 1/2); H NMR

CH₂Cl₂,

EDC^ꢃHCl, 1-[3-(dimethylamino)-propyl]-3-ethylcarbodii-

mide hydrochloride; N-hydroxy-

HOBt^ꢃH₂O,

dichloromethane;

CHCl₃,

chloroform;

(200 MHz, DMSO-d₆) d 8.67 (brd, J=8.0 Hz, 1H),

7.92–7.87 (m, 2H), 7.52–7.21 (m, 8H), 5.07 (m, 1H), 4.91

(brt, J=8.7 Hz, 1H), 3.78–3.60 (m, 2H).

benzotriazole hydrate; m-CPBA, meta-chloroperbenzoic

acid; DEAD, diethyl azodicarboxylate; TBDMSCl, tert-

butylchlorodimethylsilane; MOMCl, chloromethyl-

methyl ether; DBU, 1,8-diazabicyclo[5.4.0]undec-7-ene;

TBAF, tetrabutylammonium ﬂuoride.

N-[(1R)-2-Hydroxy-1-phenylethyl]-2,2-dimethyloctanamide

(60j). 2,2-Dimethyloctanoyl chloride was prepared by

the conventional method.²⁸73% yield; TLC R_f=0.64

1

(n-hexane/EtOAc, 1/2); H NMR (200 MHz, CDCl₃) d

7.400–7.25 (m, 5H), 6.28 (brd, J=6.6 Hz, 1H), 5.05 (m,

1H), 3.88 (brd, J=5.2 Hz, 2H), 1.51 (m, 2H), 1.24 (m,

8H), 1.96 (s, 6H), 0.86 (brt, J=6.6 Hz, 3H).

N-[(1R)-2-Hydroxy-1-phenylethyl]octanamide (60e). To

a stirred mixture of (2R)-2-amino-2-phenylethanol (4.93

g, 36 mmol) in dioxane (180 mL) and 0.5 M NaHCO₃

aq (220 mL) was added dropwise a solution of octanoyl

chloride (5.85 g, 36 mmol) in dioxane (20 mL) at 0 ^ꢁC.

After the reaction mixture was stirred for 2 h at that

temperature, it was diluted with EtOAc. The organic

layer was successively washed with 1 M HCl, 1 M

NaOH and brine before being dried over MgSO₄.

Removal of the solvent byevaporation aﬀorded a white

solid, which was washed with Et₂O/n-hexane and dried

under reduced pressure to obtain 60e as a white powder

(8.0 g, 85%): TLC R_f=0.21 (n-hexane/EtOAc, 1/2); MS

(MALDI-TOF, Pos.) m/z 286 (M+Na)⁺, 264

(M+H)⁺; IR (KBr) 3316, 1647, 1541, 1466, 1418, 1039,

N-[(1R)-2-Hydroxy-1-phenylethyl]-2-trimethyleneoctan-

amide (60k). 2-Trimethyleneoctanoyl chloride was pre-

29

pared bythe conventional method.

65% yield; TLC

1

R_f=0.64 (n-hexane/EtOAc, 1/2); H NMR (200 MHz,

CDCl₃) d 7.40–7.25 (m, 5H), 6.15 (brd, J=6.6 Hz, 1H),

5.06 (ddd, J=7.4, 5.0, 5.0 Hz, 1H), 3.90 (brs, 2H), 2.82

(brs, 1H), 2.45–2.30 (m, 2H), 1.95–1.70 (m, 6H), 1.40–

1.10 (m, 8H), 0.86 (brt, J=6.6 Hz, 3H).

N-[(1R)-2-Hydroxy-1-phenylethyl]-2-methyloctanamide

(60l₁and 60l₂). 2-Methlyoctanoyl chloride was prepared

bythe conventional method. ³⁰Puriﬁcation was performed

bycolumn chromatographyon silica gel (FL60D, n-hex-

ane/EtOAc, 3/1–2/1) to aﬀord 60l₁and 60l₂. 60l₁(less

polar, 32% yield): TLC R_f=0.64 (n-hexane/EtOAc, 1/2);

¹H NMR (200 MHz, CDCl₃) d 7.40–7.25 (m, 5H), 6.13

(brd, J=6.6 Hz, 1H), 5.06 (ddd, J=7.4, 5.0, 5.0 Hz, 1H),

3.90 (brs, 2H), 2.82 (brs, 1H), 2.35–2.15 (m, 1H), 1.80–1.20

(m, 10H), 1.17 (d, J=6.6 Hz, 3H), 0.86 (brt, J=6.6 Hz,

3H). 60l₂(more polar, 28% yield): TLC R_f=0.55 (n-hex-

ane/EtOAc, 1/2); ¹H NMR (200 MHz, CDCl₃) d 7.40–7.25

(m, 5H), 6.13 (brd, J=6.6 Hz, 1H), 5.06 (ddd, J=7.4, 5.0,

5.0 Hz, 1H), 3.90 (brd, J=5.0 Hz, 2H), 2.80 (brs, 1H),

2.35–2.17 (m, 1H), 1.70–1.10 (m, 10H), 1.15 (d, J=6.6 Hz,

3H), 0.87 (brt, J=6.6 Hz, 3H)

1

702 cm^ꢀ1; H NMR (200 MHz, CDCl₃) d 7.40–7.25 (m,

5H), 6.15 (d, J=6.2 Hz, 1H), 5.07 (m, 1H), 3.88 (dd,

J=6.6, 4.6 Hz, 2H), 2.81 (brt, J=6.0 Hz, 1H), 2.24 (brt,

J=7.6 Hz, 2H), 1.66 (m, 2H), 1.40–1.20 (m, 8H), 0.87

(brt, J=6.6 Hz, 3H).

Preparation of 60a–d, g, j, k, l_{1, 2}and m. The following

compounds were prepared from (2R)-2-amino-2-phenyl-

ethanol according to the same procedure as described

for the preparation of 60e from (2R)-2-amino-2-phenyl-

ethanol.

N-[(1R)-2-Hydroxy-1-phenylethyl]acetamide (60a). Yield,

1

88%; TLC R_f=0.20 (CH₂Cl₂/MeOH, 15/1); H NMR

(200 MHz, CDCl₃) d 7.40–7.25 (m, 5H), 6.28 (brs, 1H),

5.06 (m, 1H), 3.87 (brs, 2H), 2.86 (brs 1H), 2.04 (s, 3H).

Hexyl (1R)-2-hydroxy-1-phenylethylcarbamate (60m).

1

Yield, 99%; TLC R_f=0.47 (n-hexane/EtOAc, 2/1); H

NMR (200 MHz, CDCl₃) d 7.40–7.25 (m, 5H), 5.41

(brd, J=7.0 Hz, 1H), 4.82 (m, 1H), 4.05 (t, J=6.8 Hz,

2H), 3.86 (brs, 2H), 2.25 (brs, 1H), 1.58 (brs, 2H), 1.40–

1.20 (m, 6H), 0.87 (brt, J=6.6 Hz, 3H).

N-[(1R)-2-Hydroxy-1-phenylethyl]butanamide (60b). Yield,

1

93%; TLC R_f=0.28 (n-hexane/EtOAc, 1/2); H NMR

(300 MHz, CDCl₃) d 7.40–7.25 (m, 5H), 6.29 (brs, 1H),

5.05 (m, 1H), 3.85 (brd, J=5.1 Hz, 2H), 3.04 (brs, 1H),

2.20 (t, J=7.5 Hz, 2H), 1.67 (tq, J=7.5, 7.5 Hz, 2H),

0.94 (t, J=7.5 Hz, 3H).

N-[(1R)-2-Hydroxy-1-phenylethyl]-2-propylpentanamide

(60h). To a stirred solution of (2R)-2-amino-2-phenyl-

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3767

ethanol (500 mg, 3.65 mmol), 2-propylpentanoic acid

.

(0.63 mL, 4.02 mmol) and HOBt H₂O (1.18 g, 7.7

mmol) in DMF (5 mL) was added EDC HCl (840 mg,

conventional method.³³Puriﬁcation was performed by

column chromatographyon silica gel (FL60D, CH Cl₂/

2

.

MeOH, 40/1–35/1–30/1) to aﬀord 60p: 32% yield;

slightlyyellow oil; TLC R_f=0.64 (n-hexane/EtOAc, 1/

4.38 mmol) at 0 ^ꢁC and the mixture was stirred for 18 h.

The reaction mixture was diluted with EtOAc (20 mL).

The organic layer was successively washed with 1 M

HCl, saturated NaHCO₃aq and brine before being

dried over Na₂SO₄. Removal of the solvent byeva-

poration gave an oilyresidue, which was solidiﬁed by n-

hexane to aﬀord 60h (744 mg, 78%) as a white powder.

TLC R_f=0.47 (CHCl₃/MeOH, 9/1); ¹H NMR

(300 MHz, CDCl₃) d 7.42–7.22 (m, 5H), 6.11 (d, J=6.6

Hz, 1H), 5.15–5.02 (m, 1H), 4.00–3.80 (m, 2H), 2.84 (t,

J=6.0 Hz, 1H), 2.20–2.00 (m, 1H), 1.70–1.50 (m, 2H),

1.50–1.20 (m, 6H), 0.92 (t, J=7.2 Hz, 3H), 0.88 (t,

J=7.2 Hz, 3H).

1

2); H NMR (200 MHz, CDCl₃) d 7.40–7.25 (m, 5H),

6.39 (brd, J=6.6 Hz, 1H), 5.06 (ddd, J=7.4, 5.0, 5.0

Hz, 1H), 3.86 (brt, J=5.0 Hz, 1H), 3.45 (q, J=7.0,

2H), 3.43 (t, J=7.0 Hz, 2H), 2.99 (brs, 1H), 2.29 (t,

J=6.8 Hz, 1H), 1.80–1.55 (m, 4H), 1.17 (brt, J=7.0

Hz, 3H).

N-[(1R)-2-Hydroxy-1-phenylethyl]-6-methoxyhexanamide

(60q). 6-Methoxyhexanoic acid was prepared by the

conventional method.³⁴Puriﬁcation was performed by

column chromatographyon silica gel (Merck 7734,

CHCl₃/MeOH, 9/1) to aﬀord 60q: 56% yield; TLC

R_f=0.42 (CHCl₃/MeOH, 9/1); ¹H NMR (200 MHz,

CDCl₃) d 7.42–7.22 (m, 5H), 6.19 (d, J=7.4 Hz, 1H),

5.07 (td, J=7.4, 5.0 Hz, 1H), 3.88 (t, J=5.0 Hz, 2H),

3.37 (t, J=6.4 Hz, 2H), 3.32 (s, 3H), 2.90–2.80 (m, 1H),

2.26 (t, J=7.2 Hz, 2H), 1.80–1.50 (m, 4H), 1.50–1.30

(m, 2H).

Preparation of 60f₁, i, n, o–q, and t. The following

compounds were prepared according to the same pro-

cedure as described for the preparation of 60h from

(2R)-2-amino-2-phenylethanol.

N-[(1R)-2-Hydroxy-1-phenylethyl]undec-10-enamide (60f₁).

1

Yield, 70%; TLC R_f=0.13 (n-hexane/EtOAc, 1/1); H

N-[(1R)-2-Hydroxy-1-phenylethyl]-2-(pentylthio)acetamide

(60t). Yield, 85%; TLC R_f=0.12 (n-hexane/EtOAc, 1/

1); ¹H NMR (300 MHz, CDCl₃) d 7.56 (brd, J=6.9 Hz,

1H), 7.40–7.30 (m, 5H), 5.09 (ddd, J=7.8, 4.8, 4.8 Hz,

1H), 3.90 (brd, J=4.8 Hz, 2H), 3.30 (d, J=16.8 Hz,

1H), 3.23 (d, J=16.8 Hz, 1H), 2.53 (m, 1H), 2.53 (brt,

J=6.9 Hz, 2H), 1.65–1.50 (m, 2H), 1.40–1.20 (m, 4H),

0.87 (brt, J=7.2 Hz, 3H).

NMR (200 MHz, CDCl₃) ꢀ 7.40–7.25 (m, 5H), 6.17 (d,

J=7.0 Hz, 1H), 5.80 (dddd, J=17.5, 10.4, 5.6, 5.6 Hz,

1H), 5.10–4.90 (m, 5H), 3.88 (m, 2H), 3.30 (m, 1H), 2.23

(brt, J=7.6 Hz, 2H), 2.10–1.96 (m, 2H), 1.70–1.50 (m,

2H), 1.40–1.20 (m, 10H).

N-[(1R)-2-Hydroxy-1-phenylethyl]-5-phenylpentanamide

(60i). Yield, 53%; TLC R_f=0.35 (CH₂Cl₂/MeOH, 19/

1); H NMR (200 MHz, CDCl₃) d 7.40–7.10 (m, 10H),

N-Hexyl-N⁰-[(1R)-2-hydroxy-1-phenylethyl]urea (60r).

To a stirred solution of (2R)-2-amino-2-phenylethanol

(960 mg, 7 mmol) in CH₂Cl₂(20 mL) was added drop-

wise hexyl isocyanate (1.0 mL, 6.86 mmol) at 0 ^ꢁC and

the stirring was continued for 1 h at room temperature.

The reaction mixture was diluted with CH₂Cl₂. The

organic layer was successively washed with 1 M HCl

and brine before being dried over MgSO₄. Removal of

the solvent byevaporation gave a solid, which was

washed with Et₂O and dried under reduced pressure to

aﬀord 60r: 50% yield; TLC R_f=0.30 (CH₂Cl₂/MeOH,

19/1); ¹H NMR (300 MHz, CDCl₃) d 7.45–7.25 (m, 5H),

5.75 (brs, 1H), 4.72 (brt, J=5.0 Hz, 1H), 3.78 (m, 2H),

3.11 (m, 2H), 2.75 (brs, 1H), 1.50–1.10 (m, 8H), 0.85 (t,

J=7.0 Hz, 3H).

1

6.13 (brd, J=5.2 Hz, 1H), 5.05 (m, 1H), 3.86 (d, J=4.8

Hz, 2H), 2.62 (t, J=7.2 Hz, 2H), 2.25 (t, J=7.2 Hz,

2H), 1.80–1.55 (m, 4H).

N-[(1R)-2-Hydroxy-1-phenylethyl]-2-(pentyloxy)acetamide

(60n). (Pentyloxy)acetic acid was prepared by the con-

ventional method.³¹Puriﬁcation was performed bycol-

umn chromatographyon silica gel (FL60D, n-hexane/

EtOAc, 2/1–1/1–1/2) to aﬀord 60n: 50% yield; TLC

1

R_f=0.64 (n-hexane/EtOAc, 1/2); H NMR (200 MHz,

CDCl₃) d 7.45–7.25 (m, 5H), 7.21 (brd, J=6.2 Hz, 1H),

5.10 (ddd, J=7.4, 5.2, 5.2 Hz, 1H), 4.01 (d, J=15.0 Hz,

1H), 3.93 (d, J=15.0, 1H), 3.93–3.87 (m, 2H), 3.55–3.48

(m, 2H), 2.82 (brt, J=6.0 Hz, 1H), 1.70–1.50 (m, 2H),

1.40–1.20 (m, 4H), 0.89 (brt, J=7.2 Hz, 3H).

N-[(1R)-2-Hydroxy-1-phenylethyl]octane-1-sulfonamide

(60s). To a stirred mixture of (2R)-2-amino-2-phenyl-

ethanol (960 mg, 7 mmol) and Et₃N (2.0 mL, 14 mmol)

in CH₂Cl₂(20 mL) was added chlorotrimethylsilane

(0.90. mL, 7 mmol) at ꢀ78 ^ꢁC. Stirring was continued at

that temperature for 3 h and at 0 ^ꢁC for 1 h. To the

mixture was added dropwise 1-octanesulfonyl chloride

(1.4 mL, 7 mmol) at ꢀ78 ^ꢁC. The reaction mixture was

then stirred at that temperature for 2 h and at 0 ^ꢁC for

1.5 h. The reaction mixture was diluted with EtOAc

(100 mL) and the organic layer was successively washed

with 1 M HCl, satd NaHCO₃aq and brine before being

dried over MgSO₄. Removal of the solvent byevapora-

tion gave an oilyresidue, puriﬁcation of which was per-

formed bycolumn chromatographyon silica gel (Merck

N-[(1R)-2-Hydroxy-1-phenylethyl]-4-propoxybutanamide

(60o). 4-Propoxybutanoic acid was prepared by the

conventional method.³²Puriﬁcation was performed by

column chromatographyon silica gel (Merck 7734,

CHCl₃/MeOH, 9/1) to aﬀord 60o: 58% yield; a slightly

yellow oil: TLC R_f=0.40 (CHCl₃/MeOH, 9/1); ¹H

NMR (200 MHz, CDCl₃) d 7.42–7.22 (m, 5H), 6.55 (d,

J=7.0 Hz, 1H), 5.06 (td, J=7.0, 5.0 Hz, 1H), 3.87 (t,

J=5.0 Hz, 2H), 3.45 (t, J=6.0 Hz, 2H), 3.39–3.28 (m,

2H), 3.00 (t, J=5.8 Hz, 1H), 2.37 (t, J=7.2 Hz, 2H), 2.05–

1.80 (m, 2H), 1.65–1.40 (m, 2H), 0.88 (t, J=7.2 Hz, 3H).

5-Ethoxy-N-[(1R)-2-hydroxy-1-phenylethyl]pentanamide

(60p). 5-Ethyloxypentanoic acid was prepared by the

3768

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

7734, n-hexane/EtOAc, 2/1–3/2) to aﬀord 60s: 78% yield;

TLC R_f=0.50 (CH₂Cl₂/MeOH, 19/1); ¹H NMR

(200 MHz, CDCl₃) d 7.45–7.30 (m, 5H), 5.33 (d, J=7.0

Hz, 1H), 4.60 (m, 1H), 3.89 (dd, J=11.4, 4.4 Hz, 1H), 3.77

(dd, J=11.4, 7.0 Hz, 1H), 2.81 (ddd, J=13.8, 10.0, 6.2 Hz,

1H), 2.66 (ddd, J=13.8, 9.6, 6.0 Hz, 1H), 1.80–1.50 (m,

2H), 1.40–1.00 (m, 10H), 0.87 (t, J=7.0 Hz, 3H).

7.40–7.20 (m, 15H), 6.69 (brd, J=7.6 Hz, 1H), 5.20 (m,

1H), 4.97 (d, J=8.4 Hz, 2H), 4.94 (dd, J=11.8, 8.4 Hz,

1H), 4.87 (dd, J=11.8, 8.4 Hz, 1H), 4.18 (dd, J=8.4,

5.2 Hz, 2H), 1.96 (s, 3H). The title compound 3 was

prepared from 61a according to the same procedure as

described for the preparation of 1 from 61e: 78% yield;

oﬀ-white powder; TLC R_f=0.23 (CHCl₃/MeOH/H₂O,

65/35/8); IR (KBr) 3404, 1656, 1629, 1558, 1455, 1378,

1306, 1100 cm^ꢀ1; ¹H NMR (300 MHz, CD₃OD) d 7.40–

7.15 (m, 5H), 4.85 (m, 1H), 3.96 (m, 2H), 2.00 (s, 3H);

MS (FAB, Pos.) m/z 326 (M+Na)⁺, 304 (M+H)⁺;

General method A. (2R)-2-(Octanoylamino)-2-phenyl-

ethyl disodium phosphate (1). To a stirred solution of

60e (7.89 g, 30 mmol) in THF (200 mL) was added

dropwise n-butyllithium in n-hexane (1.53 M, 40 mL, 61

mmol) at ꢀ78 ^ꢁC under an argon atmosphere. To the

resulting mixture was added dropwise a solution of

dibenzlyphosphorochloridate³⁵in THF (1 M, 90 mL, 90

mmol) at ꢀ60 ^ꢁC and the stirring was continued for 1 h

at ꢀ78 ^ꢁC. The reaction was quenched with 1 M NaOH

(100 mL). The resulting mixture was warmed to room

temperature before being diluted with EtOAc (100 mL).

After the organic layer was successively washed with

1 M NaOH, saturated NaHCO₃aq and brine, it was

dried over MgSO₄. Removal of the solvent byevapora-

tion gave 61e, which was used for the next reaction

without further puriﬁcation. A mixture of 61e in MeOH

(200 mL) and 10% Pd–C (1 g) was stirred at room

temperature under an atmospheric pressure of hydrogen

for 2 h. Removal of the catalyst by ﬁltration through a

pad of Celite followed byevaporation aﬀorded an oily

residue which was dissolved in CH₂Cl₂(50 mL) and

extracted with 2 M NaOH (50 mL). The aqueous layer

was washed with CH₂Cl₂(50 mLꢄ2), acidiﬁed with 2 M

HCl (100 mL) and extracted with EtOAc (100 mL). The

organic layer was washed with brine, dried over Na₂SO₄

and concentrated to give 62e (4.67 g, 45% in 2 steps):

¹H NMR (200 MHz, CD₃OD) d 7.40–7.20 (m, 5H), 5.18

(brdd, J=7.2, 5.6 Hz, 1H), 4.15–4.07 (m, 2H), 2.25 (brt,

J=7.4 Hz, 2H), 1.70–1.50 (m, 2H), 1.40–1.20 (m, 8H),

0.88 (brt, J=6.6 Hz, 3H). To a stirred solution of 62e

(4.67 g, 13.6 mmol) in EtOH (200 mL) was added 1 M

NaOH (27.2 mL, 27.2 mmol) at 0 ^ꢁC. Removal of the

solvent byevaporation followed bythe dissolution of

the residue in EtOH was repeated several times to

remove H₂O azeotropically. Next addition of Et₂O fol-

lowed byevaporation aﬀorded 1 as an oﬀ-white powder

(5.18 g, 98%): TLC R_f=0.17 (CHCl₃/MeOH/H₂O, 65/

35/4); IR (KBr) 3291, 1636, 1547, 1455, 1378, 1378,

HRMS

(MALDI-TOF,

Pos.)

calcd

for

C₁₀H₁₂NO₅P^ꢃ2Na+H⁺: 304.0327; found: 304.0334.

(2R)-2-Hexanoylamino-2-phenylethyl disodium phos-

phate (5). Compound 61c was obtained from 60c

according to the same procedure as described for the

preparation of 61e from 60e and was puriﬁed bycolumn

chromatographyon silica gel (Merck 7734, n-hexane/

EtOAc, 2/1–3/2–1/1) to aﬀord 61c: 58% yield; TLC

1

R_f=0.51 (n-hexane/EtOAc, 1/1); H NMR (300 MHz,

CDCl₃) d 7.40–7.20 (m, 15H), 6.65 (brd, J=7.5 Hz,

1H), 5.22 (m, 1H), 4.97 (d, J=8.4 Hz, 2H), 4.94 (dd,

J=11.7, 8.4 Hz, 1H), 4.88 (dd, J=11.7, 8.4 Hz, 1H),

4.21–4.16 (m, 2H), 2.17 (t, J=7.5 Hz, 2H), 1.65–1.55

(m, 2H), 1.40–1.20 (m, 4H), 0.86 (t, J=6.9 Hz, 3H). The

title compound 5 was prepared from 61c according to

the same procedure as described for the preparation of 1

from 61e: 83% yield; oﬀ-white powder; TLC R_f=0.15

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3407, 1636,

1

1549, 1455, 1092, 984, 701 cm^ꢀ1; H NMR (200 MHz,

CDCl₃) d 7.40–7.10 (m, 5H), 4.90 (m, 1H), 3.95 (m,

2H), 2.33 (ddd, J=14.0, 7.2, 7.2 Hz, 1H), 2.23 (ddd,

J=14.0, 7.2, 7.2 Hz, 1H), 1.60 (m, 2H), 1.31 (m, 4H),

0.89 (brt, J=7.2 Hz, 3H); MS (FAB, Pos.) m/z 382

(M+Na)⁺, 360 (M+H)⁺; HRMS (MALDI-TOF,

+

.

Pos.) calcd for C₁₄H₂₀NO₅P 2Na+H : 360.0944;

found; 360.0953.

(2R)-2-Heptanoylamino-2-phenylethyl disodium phos-

phate (6). Compound 61d was obtained from 60d

according to the same procedure as described for the

preparation of 61e from 60e and was puriﬁed bycolumn

chromatographyon silica gel (Merck 7734, n-hexane/

EtOAc, 2/1–3/2–1/1) to aﬀord 61d: 55% yield; TLC

1

R_f=0.54 (n-hexane/EtOAc, 1/1); H NMR (300 MHz,

1

1095, 987, 700 cm^ꢀ1; H NMR (200 MHz, CD₃OD) d

CDCl₃) d 7.40–7.20 (m, 15H), 6.65 (brd, J=7.8 Hz,

1H), 5.22 (m, 1H), 4.98 (d, J=8.4 Hz, 2H), 4.94 (dd,

J=11.7, 8.4 Hz, 1H), 4.88 (dd, J=11.7, 8.4 Hz, 1H),

4.21–4.16 (m, 2H), 2.17 (t, J=7.5 Hz, 2H), 1.65–1.55

(m, 2H), 1.35–1.20 (m, 6H), 0.86 (brt, J=6.6 Hz, 3H).

The title compound 6 was prepared from 61d according

to the same procedure as described for the preparation

of 1 from 61e: 75% yield; oﬀ-white powder; TLC

R_f=0.16 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3304,

7.36–7.13 (m, 5H), 4.85 (m, 1H), 3.95 (m, 2H), 2.35 (dd,

J=13.8, 8.0 Hz, 1H), 2.21 (dd, J=13.8, 7.0 Hz, 1H),

1.59 (m, 2H), 1.40–1.20 (m, 8H), 0.88 (brt, J=7.6 Hz,

3H); MS (FAB, Pos.) m/z 410 (M+Na)⁺, 388

(M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd for

+

.

C₁₆H₂₄NO₅P 2Na+H : 388.1266; found: 388.1222;

optical rotation for free acid form [a]²_D⁵ꢀ41.4 (c 1.17,

MeOH).

1636, 1548, 1455, 1092 cm^ꢀ^{1 1}H NMR (200 MHz,

;

(2R)-2-Acetylamino-2-phenylethyl disodium phosphate

(3). Compound 61a was obtained from 60a according

to the same procedure as described for the preparation

of 61e from 60e and was puriﬁed bycolumn chroma-

tographyon silica gel (Merck 7734, CH ₂Cl₂/MeOH, 50/

1–45/1–40/1) to aﬀord 61a: 39% yield; TLC R_f=0.40

CD₃OD) d 7.40–7.10 (m, 5H), 4.90 (m, 1H), 3.95 (m,

2H), 2.33 (ddd, J=14.0, 7.2, 7.2 Hz, 1H), 2.23 (ddd,

J=14.0, 7.2, 7.2 Hz, 1H), 1.60 (m, 2H), 1.31 (m, 6H),

0.88 (brt, J=7.2 Hz, 3H); MS (FAB, Pos.) m/z 396

(M+Na)⁺, 374 (M+H)⁺; HRMS (MALDI-TOF,

+

.

Pos.) calcd for C₁₅H₂₂NO₅P 2Na+H : 374.1109;

found: 374.1113.

1

(CH₂Cl₂/MeOH, 15/1); H NMR (200 MHz, CDCl₃) d

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3769

(2R)-2-Phenyl-2-undecanoylaminoethyl disodium phos-

phate (7). Compound 61f₁was obtained from 60f₁

according to the same procedure as described for the

preparation of 61e from 60e: 57% yield; The product

was used for the next reaction without further puriﬁca-

tion. The title compound 7 was prepared from 61f₁

according to the same procedure as described for the

preparation of 1 from 61e: 87% yield; oﬀ-white powder;

TLC R_f=0.27 (CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr)

3300, 1631, 1544, 1466, 1103 cm^ꢀ1; ¹H NMR (200 MHz,

CD₃OD) d 7.36–7.16 (m, 5H), 4.80 (m, 1H), 3.96 (m,

2H), 2.29 (m, 2H), 1.58 (m, 2H), 1.40–1.15 (m, 14H),

0.88 (brt, J=6.4 Hz, 3H); MS (FAB, Pos.) m/z 452

Hz, 2H), 2.19 (t, J=7.0 Hz, 2H), 1.80–1.55 (m, 4H). The

title compound 10 was prepared from 61i according to

the same procedure as described for the preparation of 1

from 61e: 92% yield; colorless powder; TLC R_f=0.21

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3316, 1637,

1

1544, 1496, 1454, 1094, 986 cm^ꢀ1; H NMR (300 MHz,

CD₃OD) d 7.32 (d, J=7.5 Hz, 2H), 7.25–7.10 (m, 8H),

4.85 (m, 1H), 3.95 (m, 2H), 2.59 (brt, J=5.7 Hz, 2H),

2.38–2.25 (m, 2H), 1.62 (m, 4H); MS (FAB, Pos.) m/z

444 (M+Na)⁺, 422 (M+H)⁺; HRMS (MALDI-TOF,

+

.

Pos.) calcd for C₁₉H₂₂NO₅P 2Na+H : 422.1109; found:

422.1141.

(M+Na)⁺, 430 (M+H)⁺; HRMS (MALDI-TOF,

(2R)-2-(2,2-Dimethylheptanoylamino)-2-phenylethyl diso-

dium phosphate (11). Yield, 60%; colorless powder;

TLC R_f=0.18 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr)

3385, 1632, 1523, 1456, 1102 cm^ꢀ1; ¹H NMR (200 MHz,

CD₃OD) d 7.36–7.14 (m, 5H), 4.85 (m, 1H), 3.97 (brt,

J=6.2 Hz, 2H), 1.53–1.46 (m, 2H), 1.40–1.00 (m, 8H),

1.25 (s, 3H), 1.14 (s, 3H), 0.86 (brt, J=6.6 Hz, 3H); MS

+

.

Pos.) calcd for C₁₉H₃₀NO₅P 2Na+H : 430.1735;

found: 430.1686.

(2R)-2-Phenyl-2-phenoxyaminoethyl disodium phosphate

(8). Compound 61g was obtained from 60g according

to the same procedure as described for the preparation

of 61e from 60e and was used for the next reaction

without further puriﬁcation. Compound 62g was

obtained from 61g according to the same procedure as

described for the preparation of 62e from 61e and was

puriﬁed bycolumn chromatographyon silica gel

(Merck 7734, CHCl₃/MeOH/NH₄OH, 65/25/1–CHCl₃/

MeOH/H₂O, 65/25/4). The fraction including the

desired product was collected and evaporated to give an

oilyresidue, which was dissolved in EtOAc and washed

with 1 M HCl. The organic layer was washed with brine,

dried over Na₂SO₄and concentrated to give 62g (35%

yield in 2 steps). The title compound 8 was prepared

from 62g with as 92% yield according to the same pro-

cedure as described for the preparation of 1 from 62e:

oﬀ-white powder; TLC R_f=0.21 (CHCl₃/MeOH/H₂O,

65/35/8); IR (KBr) 3364, 1629, 1579, 1529, 1492, 1098

cm^ꢀ1; ¹H NMR (200 MHz, CD₃OD) d 8.00 (dd, J=8.0,

2.0 Hz, 2H), 7.50–7.39 (m, 5H), 7.32–7.15 (m, 3H), 5.10

(brt, J=8.4 Hz, 1H), 4.08 (m, 2H); MS (FAB, Pos.) m/z

(FAB, Pos.) m/z 416 (M+H)⁺; HRMS (MALDI-TOF,

+

.

Pos.) calcd for C₁₈H₂₈NO₅P 2Na+H : 416.1579;

found: 416.1578.

(2R) - 2 - Phenyl - 2 - (2 - trimethyleneheptanoylamino)ethyl

disodium phosphate (12). Yield, 43%; colorless powder;

TLC R_f=0.18 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr)

;

3376, 1629, 1522, 1106 cm^ꢀ^{1 1}H NMR (200 MHz,

CD₃OD) d 7.38–7.16 (m, 5H), 4.90 (m, 1H), 3.97 (brt,

J=6.2 Hz, 2H), 2.60–2.30 (m, 2H), 2.05–1.65 (m, 6H),

1.35–1.00 (m, 8H), 0.86 (brt, J=6.6 Hz, 3H); MS (FAB,

Pos.) m/z 450 (M+Na)⁺, 428 (M+H)⁺; HRMS

+

.

(MALDI-TOF, Pos.) calcd for C₁₉H₂₈NO₅P 2Na+H :

428.1579; found: 428.1614.

(2R)-2-(2-Methyloctanoylamino)-2-phenylethyl disodium

phosphate (13a). Yield, 33%; colorless powder; TLC

R_f=0.25 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3329,

;

1625, 1533, 1466, 1112 cm^ꢀ^{1 1}H NMR (300 MHz,

366 (M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd for

CD₃OD) d 7.34 (brd, J=6.9 Hz, 2H), 7.25 (brt, J=7.2

Hz, 2H), 7.16 (m, 1H), 4.85 (m, 1H), 4.03–3.88 (m, 2H),

2.44 (m, 1H), 1.60 (m, 1H), 1.40–1.20 (m, 9H), 1.10 (d,

J=6.9 Hz, 3H), 0.88 (brt, J=6.6 Hz, 3H); MS (FAB,

+

.

C₁₅H₁₄NO₅P 2Na+H : 366.0483; found: 366.0470.

(2R)-2-Phenyl-2-(2-propylpentanoylamino)ethyl disodium

phosphate (9). White powder; TLC R_f=0.31 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3304, 2957, 2933, 2873,

1628, 1540, 1496, 1456, 1380, 1258, 1217, 1106 cm^ꢀ1; ¹H

NMR (200 MHz, CD₃OD) d 7.40–7.10 (m, 5H), 5.00–

4.78 (m, 1H), 4.05–3.90 (m, 2H), 2.50–2.30 (m, 1H),

1.70–1.10 (m, 8H), 0.90 (t, J=6.8 Hz, 3H), 0.87 (t,

J=6.8 Hz, 3H); MS (FAB, Pos.) m/z 410 (M+Na)⁺,

388 (M+H)⁺, 366; HRMS (MALDI-TOF, Pos.) calcd

Pos.) m/z 402 (M+H)⁺; HRMS (MALDI-TOF, Pos.)

+

.

calcd for C₁₇H₂₆NO₅P 2Na+H : 402.1422; found:

402.1420.

(2R)-2-(2-Methyloctanoylamino)-2-phenylethyl disodium

phosphate (13b). Yield, 91%; pinkish powder; TLC

R_f=0.20 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3315,

1631, 1540, 1455, 1377, 1108 cm^ꢀ1; ¹H NMR (300 MHz,

CD₃OD) d 7.36 (brd, J=7.2 Hz, 2H), 7.25 (brt, J=7.2

Hz, 2H), 7.17 (m, 1H), 4.85 (m, 1H), 4.00–3.90 (m, 2H),

2.48 (m, 1H), 1.55 (m, 1H), 1.40–1.15 (m, 9H), 1.08 (d,

J=6.6 Hz, 3H), 0.88 (brt, J=6.6 Hz, 3H); MS (FAB,

+

.

for C₁₆H₂₄NO₅P 2Na+H : 388.1266; found: 388.1302.

(2R)-2-Phenyl-2-(5-phenylpentanoylamino)ethyl disodium

phosphate (10). Compound 61i was obtained from 60i

according to the same procedure as described for the

preparation of 61e from 60e and was puriﬁed bycolumn

chromatographyon silica gel (Merck 7734, CH ₂Cl₂/

MeOH, 50/1) to aﬀord 61i: 54% yield; ¹H NMR

(200 MHz, CDCl₃) d 7.40–7.10 (m, 20H), 6.65 (brd,

J=7.6 Hz, 1H), 5.21 (m, 1H), 4.95 (d, J=8.4 Hz, 2H),

4.93 (dd, J=11.6, 8.4 Hz, 1H), 4.86 (dd, J=11.6, 8.4

Hz, 1H), 4.18 (dd, J=8.6, 5.0 Hz, 2H), 2.58 (t, J=7.0

Pos.) m/z 424 (M+Na)⁺, 402 (M+H)⁺; HRMS

+

.

(MALDI-TOF, Pos.) calcd for C₁₇H₂₆NO₅P 2Na+H :

402.1422; found: 402.1461.

(2R)-2-(Hexanoyloxycarbonylamino)-2-phenylethyl diso-

dium phosphate (14). Yield, 52%; colorless powder;

TLC R_f=0.16 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr)

;

3409, 1696, 1455, 1421, 1341, 1104 cm^ꢀ^{1 1}H NMR

3770

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

(200 MHz, CD₃OD) d 7.36–7.14 (m, 5H), 4.70 (m, 1H),

3.97–3.78 (m, 4H), 1.65–1.10 (m, 8H), 0.88 (m, 3H); MS

(FAB, Pos.) m/z 412 (M+Na)⁺, 390 (M+H)⁺; HRMS

(MALDI-TOF, Pos.) calcd for C₁₅H₂₂NO₆P 2Na+H :

390.1058; found: 390.1067.

(2R)-2-phenyl-2-(octylsulfonylamino)ethyl disodium phos-

phate (20). Yield, 25%; colorless powder; TLC

R_f=0.25 (CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 1656,

1455, 1303, 1145 cm^ꢀ1; ¹H NMR (200 MHz, CD₃OD) d

7.45–7.23 (m, 5H), 4.57 (dd, J=7.4, 5.6 Hz, 1H), 3.92

(m, 2H), 2.77 (m, 2H), 1.65 (m, 2H), 1.40–1.10 (m,

10H), 0.88 (brt, J=7.0 Hz, 3H); MS (FAB, Pos.) m/z

+

.

(2R)-2-Phenyl-2-(pentyloxyethanoylamino)ethyl disodium

phosphate (15). Yield, 80%; colorless powder; TLC

R_f=0.18 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3405,

460 (M+Na)⁺, 438 (M+H)⁺; HRMS (MALDI-TOF,

+

.

Pos.) calcd for C₁₆H₂₆NO₆P 2Na+H : 438.1092;

found: 438.1079.

1662, 1542, 1454, 1102, 987, 699 cm^ꢀ1

;

¹H NMR

(300 MHz, CD₃OD) d 7.36 (d, J=7.5 Hz, 2H), 7.30–

7.19 (m, 3H), 5.03 (dd, J=8.1, 7.2 Hz, 1H), 4.03 (d,

J=16.5 Hz, 1H), 4.00 (m, 2H), 3.95 (d, J=16.5 Hz,

1H), 3.50 (t, J=6.9 Hz, 2H), 1.70–1.55 (m, 2H), 1.50–

1.30 (m, 4H), 0.90 (brt, J=6.0 Hz, 3H); MS (FAB, Pos.)

m/z 412 (M+Na)⁺, 390 (M+H)⁺; HRMS (MALDI-

General method B

(2R)-2-Butanoylamino-2-phenylethyl disodium phosphate

(4). To a stirred solution of 60b (207 mg, 1 mmol) in

THF (10 mL) was added dropwise lithium diisopropy-

lamide in THF (2 M, 0.53 mL, 1.06 mmol) at ꢀ78 ^ꢁC

under an argon atmosphere. To the resulting mixture

was added tetrabenzylpirophosphate³⁶(646 mg, 1.2

mmol) in a single portion. Stirring was continued for 1 h

at ꢀ78 ^ꢁC and at 0 ^ꢁC for 30 min. The reaction was

quenched with saturated NaHCO₃aq (10 mL) and

extracted with EtOAc (50 mL). The organic layer was

washed with brine and dried over MgSO₄. Removal of

the solvent byevaporation gave an oilyresidue. The

product was puriﬁed bycolumn chromatographyon

silica gel (Merck 7734, CH₂Cl₂/MeOH, 40/1) to aﬀord

61b (343 mg, 73%): ¹H NMR (300 MHz, CDCl₃) d

7.40–7.20 (m, 15H), 6.64 (brd, J=7.5 Hz, 1H), 5.22 (m,

1H), 4.97 (d, J=8.4 Hz, 2H), 4.94 (dd, J=11.7, 8.4 Hz,

1H), 4.87 (dd, J=11.7, 8.4 Hz, 1H), 4.19 (dd, J=8.4,

5.1 Hz, 2H), 2.15 (t, J=7.5 Hz, 2H), 1.70–1.57 (m, 2H),

0.91 (t, J=7.5 Hz, 3H). The title compound 4 was

obtained from 61b according to the same procedure as

described for the preparation of 1 from 61e as an oﬀ-

white powder: TLC R_f=0.12 (CHCl₃/MeOH/H₂O, 65/

+

.

TOF, Pos.) calcd for C₁₅H₂₂NO₆P 2Na+H : 390.1058;

found: 390.1104.

(2R)-2-Phenyl-2-(propyloxybutanoylamino)ethyl disodium

phosphate (16). Yield, 87%; white powder; TLC

R_f=0.28 (CHCl₃/MeOH/H₂O, 65/25/8); IR (KBr) 3290,

2960, 2874, 1646, 1549, 1496, 1455, 1379, 1276, 1110

1

cm^ꢀ1; H NMR (200 MHz, CD₃OD) d 7.40–7.10 (m,

5H), 5.02–4.85 (m, 1H), 4.10–3.90 (m, 2H), 3.42 (t,

J=6.5 Hz, 2H), 3.36 (t, J=6.8 Hz, 2H), 2.50–2.20 (m,

2H), 1.98–1.78 (m, 2H), 1.70–1.45 (m, 2H), 0.91 (t,

J=7.5 Hz, 3H); MS (FAB, Pos.) m/z 412 (M+Na)⁺,

390 (M+H)⁺, 368; HRMS (MALDI-TOF, Pos.) calcd

+

.

for C₁₅H₂₂NO₆P 2Na+H : 390.1058; found: 390.1073.

(2R)-2-Ethyloxypentanoylamino-2-phenylethyl disodium

phosphate (17). Yield, 82%; colorless powder; TLC

R_f=0.13 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3298,

1639, 1548, 1455, 1378, 1108 cm^ꢀ1; ¹H NMR (300 MHz,

CD₃OD) d 7.36 (d, J=7.5 Hz, 2H), 7.30–7.15 (m, 3H),

4.90 (m, 1H), 3.95 (m, 2H), 3.45 (q, J=7.2 Hz, 2H), 3.41

(t, J=6.0 Hz, 2H), 2.31 (m, 2H), 1.70–1.50 (m, 4H), 1.15

(t, J=7.2 Hz, 3H); MS (FAB, Pos.) m/z 412 (M+Na)⁺,

390 (M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd for

25/4); IR (KBr) 3306, 1637, 1548, 1456, 1093 cm^ꢀ1; H

1

NMR (200 MHz, CDCl₃) d 7.40–7.15 (m, 5H), 4.85 (m,

1H), 3.96 (m, 2H), 2.26 (m, 2H), 1.63 (ddq, J=7.2, 7.2,

7.2 Hz, 2H), 0.93 (t, J=7.2 Hz, 3H); MS (FAB, Pos.) m/z

354 (M+Na)⁺, 332 (M+H)⁺; HRMS (MALDI-TOF,

.

Pos.) calcd for C₁₂H₁₆NO₅P 2Na+H : 332.0640;

found: 332.0624.

+

.

C₁₅H₂₂NO₆P 2Na+H : 390.1058; found: 390.1085.

+

(2R)-2-Methoxyhexanoylamino-2-phenylethyl disodium

phosphate (18). Yield, 57%; white powder; TLC

R_f=0.26 (CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3284,

;

2935, 2865, 1634, 1549, 1496, 1455, 1119 cm^ꢀ^{1 1}H

General method C

NMR (300 MHz, CD₃OD) d 7.40–7.14 (m, 5H), 4.98–

4.82 (m, 1H), 4.02–3.85 (m, 2H), 3.37 (t, J=6.5 Hz, 2H),

3.30 (s, 3H), 2.40–2.20 (m, 2H), 1.70–1.50 (m, 4H),

1.50–1.30 (m, 2H); MS (FAB, Pos.) m/z 412 (M+Na)⁺,

(2R)-2-(2-Pentylthioethanoylamino)-2-phenylethyl disodium

phosphate (22). To a stirred solution of 60t (281 mg, 1

mmol) in benzene (2 mL) was added NaH (60% dis-

persion in mineral oil, 80 mg, 2 mmol) at room tem-

perature under an argon atmosphere. The resulting

mixture was heated under reﬂux for 30 min. After cool-

ing, the mixture was diluted with THF (2 mL) and then

di-tert-butyl phosphorobromidate³⁷(272 mg, 1 mmol)

was added. Stirring was continued at 30 ^ꢁC for 2 h. The

reaction mixture was diluted with EtOAc. The organic

layer was washed with saturated NaHCO₃aq and brine

before being dried over MgSO₄. Removal of the solvent

byevaporation gave 61t as an oil, which was used for

the next reaction without further puriﬁcation. To a stirred

solution of 61t in benzene (5 mL) was added CF₃COOH

(1 mL) and the mixture stirred at room temperature for 15

h. Removal of the solvent byevaporation gave an oily

390 (M+H)⁺, 368; HRMS (MALDI-TOF, Pos.) calcd

+

.

for C₁₅H₂₂NO₆P 2Na+H : 390.1058; found: 390.1101.

(2R)-2-(N-Hexylureido)-2-phenylethyl disodium phosphate

(19). Yield, 63%; oﬀ-white powder; TLC R_f=0.23

(CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3323, 1648,

1560, 1454, 1100 cm^ꢀ1; ¹H NMR (200 MHz, CD₃OD) d

7.37–7.11 (m, 5H), 4.73 (dd, J=8.0, 4.0 Hz, 1H), 4.03–

3.80 (m, 2H), 3.05 (t, J=6.8 Hz, 2H), 1.50–1.20 (m, 8H),

0.88 (brt, J=7.0 Hz, 3H); MS (FAB, Pos.) m/z 411

(M+Na)⁺, 389 (M+H)⁺; HRMS (MALDI-TOF,

+

.

Pos.) calcd for C₁₅H₂₃NO₅P 2Na+H : 389.1218;

found: 389.1211.

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3771

residue, which was partitioned between CH₂Cl₂(5 mL)

and 2 M NaOH (5 mL). The aqueous layer was washed

with CH₂Cl₂(ꢄ3), acidiﬁed byadding 2 M HCl (10 mL)

and extracted with EtOAc. The organic layer was dried

over NaSO₄. Removal of the solvent byevaporation

gave 62t as an oil (74% in 2 steps), which was converted

to the disodium salt according to the same procedure as

described for preparation of 1 from 62e: 75% yield;

orange powder; TLC R_f=0.35 (CHCl₃/MeOH/H₂O, 65/

of LiBH₄(549 mg, 25 mmol) in THF (25 mL) was

added dropwise a solution of methyl (2-methoxy-

phenyl)(octanoylamino)acetate (4.0g, 12.5 mmol) in

THF (10 mL) at 0 ^ꢁC and stirring was continued for 20

h at room temperature. The reaction was quenched by

adding satd NH₄Cl aq and the mixture was diluted with

EtOAc. The organic layer was washed with brine and

dried over Na₂SO₄. Removal of the solvent byeva-

poration gave an oilyresidue, which was solidiﬁed with

n-hexane to aﬀord 64a as a white powder: 98% yield;

TLC R_f=0.52 (CHCl₃/MeOH, 9/1); ¹H NMR (200 MHz,

CDCl₃) d 7.38–7.20 (m, 2H), 7.00–6.85 (m, 2H), 6.53 (d,

J=7.6 Hz, 1H), 5.40–5.28 (m, 1H), 4.00–3.70 (m, 5H),

2.55 (t, J=5.6 Hz, 1H), 2.30–2.20 (m, 2H), 1.75–1.50 (m,

2H), 1.40–1.15 (m, 8H), 0.87 (t, J=6.6 Hz, 3H).

1

25/8); IR (KBr) 3342, 1640, 1543, 1454, 1100 cm^ꢀ1; H

NMR (300 MHz, CD₃OD) d 7.37 (brd, J=7.5 Hz, 2H),

7.26 (brt, J=7.5 Hz, 2H), 7.18 (m, 1H), 4.92 (dd,

J=8.1, 4.5 Hz, 1H), 3.97 (m, 2H), 3.36 (d, J=13.8 Hz,

1H), 3.13 (d, J=13.8 Hz, 1H), 2.52 (brt, J=7.2 Hz, 2H),

1.54 (m, 2H), 1.30 (m, 4H), 0.87 (brt, J=7.2 Hz, 3H);

MS (FAB, Pos.) m/z 428 (M+Na)⁺, 406 (M+H)⁺;

HRMS

(MALDI-TOF,

Pos.)

calcd

for

Preparation of 64b–k and 68a–e: the following com-

pounds were prepared according to essentiallythe same

procedures as described for the preparation of 64a from

63a.

C₁₅H₂₂NO₅PS^ꢃ2Na+H⁺: 406.0830; found: 406.0873.

(2R)-2-(2-Pentylsulfonylethanoylamino) -2-phenylethyl

disodium phosphate (21). To a stirred solution of 62t

(166 mg, 0.45 mmol) in MeOH (2.5 mL) was added

N-[2-Hydroxy-1-(3-methoxyphenyl)ethyl]octanamide (64b).

Yield, 94%; white powder; TLC R_f=0.53 (CHCl₃/

dropwise a solution of OXONE¹(2KHSO₅KH-

.

1

.

SO₄K₂SO₄, 311 mg, 0.505 mmol) in H₂O (2.5 mL) at

MeOH, 9/1); H NMR (200 MHz, CDCl₃) d 7.35–7.22

0 ^ꢁC. Stirring was continued at 0 ^ꢁC for 3 h and at room

temperature for 15 h. A precipitate which formed was

removed byﬁltration. The solid was washed with

MeOH. The ﬁltrate was combined and concentrated to

give an oilyresidue, which was dissolved in EtOAc. The

organic layer was washed with 1 M HCl and dried over

Na₂SO₄. Removal of the solvent byevaporation gave

(2R)-2-(2-pentylsulfonylamino)-2-phenylethyl dihydro-

gen phosphate as an oil (75%), which was converted to

the disodium salt according to the same procedure as

described for preparation of 1 from 62e: 85% yield;

orange powder; TLC R_f=0.16 (CHCl₃/MeOH/H₂O, 65/

(m, 1H), 6.95–6.80 (m, 3H), 6.13 (d, J=6.4 Hz, 1H),

5.10–4.98 (m, 1H), 3.95–3.81 (m, 2H), 3.81 (s, 3H),

2.80–2.65 (br, 1H), 2.30–2.20 (m, 2H), 1.80–1.50 (m,

2H), 1.50–1.18 (m, 8H), 0.87 (t, J=6.6 Hz, 3H).

N-[2-Hydroxy-1-(4-methoxyphenyl)ethyl]octanamide (64c).

Methyl (4-methoxylphenyl)(octanoylamino)acetate was

prepared from 63c according to the same procedure as

described for the preparation of 64a from 63a using

NaHCO₃as a base instead of pyridine and the product

was used for the next reaction without further puriﬁ-

1

cation: TLC R_f=0.30 (CH₂Cl₂/MeOH, 19/1); H NMR

25/4); IR (KBr) 3212, 1668, 1564, 1455, 1298, 1123 cm^ꢀ1

;

(300 MHz, CDCl₃) d 7.21 (d, J=9.0 Hz, 2H), 6.88 (d,

J=9.0 Hz, 2H), 6.20 (d, J=6.6 Hz, 1H), 4.99 (m, 1H),

3.83 (m, 2H), 3.79 (s, 3H), 3.40–2.80 (brs, 1H), 2.22 (t,

J=7.2 Hz, 2H), 1.63 (m, 2H), 1.40–1.20 (m, 8H), 0.87 (t,

J=6.9 Hz, 3H).

¹H NMR (300 MHz, CD₃OD) d 7.40 (brd, J=7.2 Hz,

2H), 7.30–7.15 (m, 3H), 4.91 (dd, J=8.1, 3.9 Hz, 1H),

4.05–3.90 (m, 2H), 3.17 (m, 2H), 1.78 (m, 2H), 1.45–1.30

(m, 4H), 0.89 (brt, J=7.2 Hz, 3H); MS (FAB, Pos.) m/z

438 (M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd for

+

.

C₁₅H₂₂NO₇PS 2Na+H : 438.0728; found: 438.0692.

N-[2-Hydroxy-1-(2-methylphenyl)ethyl]octanamide (64d).

Methyl (2-methylphenyl)(octanoylamino)acetate was

prepared from 63d according to the same procedure as

described for the preparation of 64a from 63a using

Et₃N as a base instead of pyridine and the product was

used for the next reaction without further puriﬁcation:

TLC R_f=0.43 (n-hexane/EtOAc, 3/1); ¹H NMR

(200 MHz, CDCl₃) d 7.23–7.16 (m, 4H), 6.30 (brd,

J=7.4 Hz, 1H), 5.81 (d, J=7.4 Hz, 1H), 3.71 (s, 3H),

2.48 (s, 3H), 2.26–2.19 (m, 2H), 1.63–1.52 (m, 2H),

1.30–1.15 (m, 8H), 0.95–0.82 (m, 3H). The title com-

pound 64d was prepared from methyl(2-methylphenyl)

(octanoylamino)acetate according to the same proce-

dure as described for the preparation of 64a from

methyl (2-methoxyphenyl)(octanoylamino)acetate. The

product was washed with Et₂O/n-hexane and used for

the next reaction without further puriﬁcation.

N-[2-Hydroxy-1-(2-methoxyphenyl)ethyl]octanamide (64a).

To a stirred suspension of 63a¹⁶(4.73 g, 24.3 mmol) in

CH₂Cl₂(32 mL) were added a solution of octanoyl

chloride (4.72 g, 29 mmol) in CH₂Cl₂(8 mL) and pyri-

dine (16 mL) at 0 ^ꢁC. Stirring was continued for 18 h at

room temperature. The reaction mixture was diluted

with EtOAc. The organic layer was successively washed

with 1 M HCl, satd NaHCO₃aq and brine before being

dried over Na₂SO₄. Removal of the solvent byeva-

poration aﬀorded a crude product which was puriﬁed

bycolumn chromatographyon silica gel (Merck 7734,

n-hexane/EtOAc, 2/1) and solidiﬁed with n-hexane to

give methyl (2-methoxyphenyl)(octanoylamino)acetate

as a slightly yellow powder: 65% yield; TLC R_f=0.21

1

(n-hexane/EtOAc, 1/1); H NMR (200 MHz, CDCl₃) d

7.40–7.25 (m, 2H), 7.00–6.85 (m, 2H), 6.54 (d, J=8.5

Hz, 1H), 5.80 (d, J=8.5 Hz, 1H), 3.84 (s, 3H), 3.68 (s,

3H), 2.30–2.15 (m, 2H), 1.70–1.50 (m, 2H), 1.40–1.15

(m, 8H), 0.86 (t, J=6.6 Hz, 3H). To a stirred suspension

N-[2-Hydroxy-1-(3-methylphenyl)ethyl]octanamide (64e).

Methyl (3-methylphenyl)(octanoylamino)acetate was

prepared from 63e according to the same procedure as

3772

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

described for the preparation of 64a from 63a using

Et₃N as a base instead of pyridine and the product was

used for the next reaction without further puriﬁcation:

TLC R_f=0.35 (n-hexane/EtOAc, 3/1); ¹H NMR

(200 MHz, CDCl₃) d 7.29–7.21 (m, 1H), 7.16–7.12 (m,

3H), 6.38 (brd, J=6.8 Hz, 1H), 3.73 (s, 3H), 2.27–2.20

(m, 2H), 1.63–1.52 (m, 2H), 1.31–1.15 (m, 8H), 0.96–

0.80 (m, 3H). The title compound 64e was prepared

from methyl (3-methylphenyl)(octanoylamino)acetate

according to the same procedure as described for the

preparation of 64a from methyl (2-methoxyphenyl)

(octanoylamino)acetate. The product was washed with

Et₂O/n-hexane and used for the next reaction without

further puriﬁcation.

N-{2-Hydroxy-1-[3-(methylthio)phenyl]ethyl}octanamide

(64k). Methyl (3-methylthiophenyl)(octanoylamino)

acetate was prepared from 63k according to the same

procedure as described for the preparation of 64a from

63a using Et₃N as a base instead of pyridine and the

product was washed with Et₂O/n-hexane: yellow pow-

der; 38% yield; TLC R_f=0.26 (n-hexane/EtOAc, 3/1).

The title compound 64k was prepared from methyl

(3-methylthiophenyl)(octanoylamino)acetate according

to the same procedure as described for the preparation

of 64a from methyl (2-methoxyphenyl)(octanoylamino)

acetate. The product was washed with Et₂O/n-hexane and

used for the next reaction without further puriﬁcation.

N-[2-Hydroxy-1-(1-naphthyl)ethyl]octanamide (68a). Ivory

powder; 73% yield; TLC R_f=0.27 (n-hexane/EtOAc,

1/3); H NMR (300 MHz, CDCl₃) d 8.08–8.05 (m, 1H),

N-[2-Hydroxy-1-(4-methylphenyl)ethyl]octanamide (64f).

1

Yield, 89%; TLC R_f=0.28 (n-hexane/EtOAc, 1/3); H

NMR (200 MHz, CDCl₃) d 7.16 (s, 4H), 6.10 (d, J=6.3

Hz, 1H), 5.05–4.99 (m, 1H), 3.88 (dd, J=11.4, 5.7 Hz,

1H), 3.83 (dd, J=11.4, 4.5 Hz, 1H), 2.33 (s, 3H), 2.25–

2.20 (m, 2H), 1.66–1.58 (m, 2H), 1.29–1.26 (m, 8H),

0.89–0.84 (m, 3H).

7.90–7.80 (m, 2H), 7.58–7.44 (m, 4H), 6.13 (d, J=6.3

Hz, 1H), 5.93–5.87 (m, 1H), 4.14–4.04 (m, 2H), 2.89

(brs, 1H), 2.37–2.17 (m, 2H), 1.70–1.60 (m, 2H), 1.28–

1.25 (m, 8H), 0.88–0.84 (m, 3H).

N-[2-Hydroxy-1-(2-naphthyl)ethyl]octanamide (68b). Ivory

powder; 77% yield; TLC R_f=0.24 (n-hexane/EtOAc,

1/3); H NMR (300 MHz, CDCl₃) d 7.84–7.73 (m, 4H),

N-[1-(2-Chlorophenyl)-2-hydroxyethyl]octanamide (64g).

1

Yield, 84%; TLC R_f=0.45 (n-hexane/EtOAc, 1/3); H

NMR (200 MHz, CDCl₃) d 7.40–7.37 (m, 1H), 7.32–

7.22 (m, 3H), 6.32 (d, J=6.3 Hz, 1H), 5.47–5.42 (m,

1H), 3.91 (d, J=4.5 Hz, 2H), 2.25 (t, J=6.6 Hz, 2H),

1.69–1.59 (m, 2H), 1.30–1.26 (m, 8H), 0.89–0.85 (m,

3H).

7.51–7.46 (m, 2H), 7.38 (dd, J=8.4 Hz, 1.8 Hz, 1H),

6.29 (d, J=6.9 Hz, 1H), 5.24–5.19 (m, 1H), 4.01–3.90

(m, 2H), 2.26 (t, J=7.2 Hz, 2H), 1.70–1.60 (m, 2H),

1.29–1.26 (m, 8H), 0.88–0.84 (m, 3H).

N-[1-(1,3-Benzodioxol-4-yl)-2-hydroxyethyl]octanamide

(68c). Oﬀ-white powder; 57% yield; TLC R_f=0.25

(n-hexane/EtOAc, 1/1); H NMR (200 MHz, CDCl₃) d

N-[1-(3-Chlorophenyl)-2-hydroxyethyl]octanamide (64h).

1

Yield, 73%; TLC R_f=0.43 (n-hexane/EtOAc, 1/3); H

NMR (300 MHz, CDCl₃) d 7.29–7.23 (m, 3H), 7.17–

7.14 (m, 2H), 6.38–6.36 (d, J=7.2 Hz, 1), 5.03–4.97 (m,

1H), 3.87–3.77 (m, 2H), 2.22 (t, J=7.2 Hz, 2H), 1.68–

1.57 (m, 2H), 1.31–1.22 (m, 8H), 0.86 (t, J=7.2 Hz, 3H).

6.90–6.75 (m, 3H), 6.35 (d, J=7.5 Hz, 1H), 6.00 (m,

2H), 5.23 (m, 1H), 4.00–3.78 (m, 2H), 2.62–2.38 (m,

1H), 2.25 (t, J=7.5 Hz, 2H), 1.78–1.45 (m, 2H), 1.40–

1.20 (m, 8H), 0.85 (m, 3H).

N-[1-(4-Chlorophenyl)-2-hydroxyethyl]octanamide (64i).

1

N-[1-(1,3-Benzodioxol-5-yl)-2-hydroxyethyl]octanamide

(68d). Methyl (1,3-dioxaindan-5-yl)(octanoylamino)

acetate was prepared from 67d according to the same

procedure as described for the preparation of 64a from

63a using Et₃N as a base instead of pyridine and the

product was washed with Et₂O/n-hexane; white powder;

82% yield; TLC R_f=0.46 (n-hexane/EtOAc, 2/1). The

title compound 68d was prepared from methyl (1,3-

dioxaindan-5-yl)(octanoylamino)acetate according to

the same procedure as described for the preparation of

64a from methyl (2-methoxyphenyl)(octanoylamino)

acetate. The product was washed with Et₂O/n-hexane and

used for the next reaction without further puriﬁcation.

Yeild, 90%; TLC R_f=0.25 (n-hexane/EtOAc, 1/3); H

NMR (300 MHz, CDCl₃) d 7.33 (d, J=8.7 Hz, 2H), 7.24

(d, J=8.7 Hz, 2H), 6.12 (d, J=6.9 Hz, 1H), 5.07–5.02

(m, 1H),3.87 (d, J=8.1 Hz,2H), 2.24 (t, J=7.5 Hz, 2H),

1.67–1.62 (m, 2H),1.28–1.25 (m, 8H), 0.89–0.85 (m, 3H).

N-[1-(3,5-Dimethoxyphenyl)-2-hydroxyethyl]octanamide

(64j). Methyl (3,5-dimethoxylphenyl)(octanoylamino)

acetate was prepared from 63j according to the same

procedure as described for the preparation of 64a from

63a using Et₃N as a base instead of pyridine and the

reaction product was washed with Et₂O/n-hexane: yel-

low powder; 50% yield; TLC R_f=0.36 (n-hexane/

1

EtOAc, 2/1); H NMR (200 MHz, CDCl₃) d 6.50 (d,

N-(2-Hydroxy-1-thien-2-ylethyl)octanamide (68e). Methyl

(octanoylamino)(thiophen-2-yl)acetate was prepared

from 67e according to the same procedure as described

for the preparation of 64a from 63a using Et₃N as a

base instead of pyridine and puriﬁed by column

chromatographyon silica gel (Merck 7734, n-hexane/

EtOAc, 4/1): 71% yield; TLC R_f=0.45 (n-hexane/

J=3.3 Hz, 2H), 6.41 (t, J=3.3 Hz, 1H), 5.51 (d, J=7.4

Hz, 1H), 3.78 (s, 3H), 3.74 (s, 3H), 2.28–2.20 (m, 2H),

1.70–1.55 (m, 2H), 1.30–1.25 (m, 8H), 0.85–0.83 (m,

3H). The title compound 64j was prepared from methyl

(3,5-dimethoxylphenyl)(octanoylamino)acetate accord-

ing to the same procedure as described for the prepara-

tion of 64a from methyl (2-methoxyphenyl)

(octanoylamino)acetate. The product was washed with

Et₂O/n-hexane and used for the next reaction without

further puriﬁcation.

1

EtOAc, 2/1); H NMR; (200 MHz, CDCl₃) d 7.26 (dd,

J=5.2, 1.2 Hz, 1H), 7.07–7.04 (m, 1H), 6.97 (dd, J=5.2,

3.8 Hz, 1H), 6.38 (brd, J=7.8 Hz, 1H), 5.89 (dd, J=7.8,

0.8 Hz, 1H), 3.79 (s, 3H), 2.27–2.21 (m, 2H), 1.71–1.55

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3773

(m, 2H), 1.37–1.22 (m, 8H), 0.92–0.82 (m, 3H). The title

compound 68e was prepared from methyl (octanoyl-

amino)(thiophen-2-yl)acetate according to the same

procedure as described for the preparation of 64a from

methyl (2-methoxyphenyl)(octanoylamino)acetate. The

product was puriﬁed bycolumn chromatographyon

silica gel (Merck 7734, n-hexane/EtOAc, 3/1).

MeOH/H₂O, 65/35/8); IR (KBr) 3259, 2925, 1625, 1557,

1

1459, 1099, 990 cm^ꢀ1; H NMR (300 MHz, CD₃OD) d

7.17–7.10 (m, 3H), 7.04–6.90 (m, 1H), 4.85 (dd, J=6.3,

4.5 Hz, 1H), 4.00–3.86 (m, 2H), 2.38–2.18 (m, 2H), 2.92

(s, 3H), 1.68–1.52 (m, 2H), 1.38–1.20 (m, 8H), 0.88 (brt,

J=6.3 Hz, 3H); MS (FAB, Pos.) m/z 402 (M+H)⁺,

380, 358; HRMS (MALDI-TOF, Pos.) calcd for free

acid form C₁₇H₂₈NO₅P+Na⁺: 380.1603; found:

380.1639.

Preparation of 23a–24c and 26: the following com-

pounds were prepared according to the same procedures

as described for the preparation of 1 from 60e (general

method A).

2-(4-Methylphenyl)-2-octanoylaminoethyl disodium phos-

phate (24c). White powder; TLC R_f=0.30 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3258, 2927, 2857, 2360,

;

1625, 1554, 1462, 1377, 1276, 1104 cm^ꢀ^{1 1}H NMR

2 - (2 - Methoxyphenyl) - 2 - octanoylaminoethyl disodium

phosphate (23a). White powder; TLC R_f=0.28 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3274, 2929, 2857, 1630,

1555, 1494, 1462, 1375, 1289, 1247, 1098, 1029 cm^ꢀ1; ¹H

NMR (300 MHz, CD₃OD) d 7.26 (d, J=7.5 Hz, 1H),

7.17 (dd, J=7.5, 7.5 Hz, 1H), 6.90 (d, J=7.5 Hz, 1H),

6.85 (dd, J=7.5, 7.5 Hz, 1H), 5.29 (dd, J=7.8, 3.6 Hz,

1H), 4.08–3.95 (m, 1H), 3.95–3.80 (m, 4H), 2.40–2.20

(m, 2H), 1.70–1.50 (m, 2H), 1.40–1.20 (m, 8H), 0.89 (t,

J=6.6 Hz, 3H); MS (FAB, Pos.) m/z 440 (M+Na)⁺,

418 (M+H)⁺, 396; HRMS (MALDI-TOF, Pos.) calcd

for C₁₇H₂₆NO₆P^ꢃ2Na+H⁺: 418.1371; found: 418.1375.

(300 MHz,CD₃OD) d 7.22 (d, J=8.1 Hz, 2H), 7.08 (d,

J=8.1 Hz, 2H), 4.90–4.85 (m, 1H), 3.99–3.86 (m, 2H),

2.36–2.17 (m, 2H), 2.27 (s, 3H), 1.67–1.52 (m, 2H),

1.38–1.24 (m,8H), 0.93–0.86 (m, 3H); MS (FAB, Pos.)

m/z 402 (M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd

+

.

for C₁₇H₂₆NO₅P 2Na+H : 402.1422; found: 402.1377.

2-(3,5-Dimethoxyphenyl)-2-octanoylaminoethyl disodium

phosphate (26). Oﬀ-white amorphous powder; TLC

R_f=0.50 (CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3291,

1609, 1549, 1464, 1206, 1155, 1102, 984 cm^ꢀ1; ¹H NMR

(300 MHz,CD₃OD) d 6.51 (d, J=2.4 Hz, 2H), 6.30 (t,

J=2.4 Hz, 1H), 4.83 (dd, J=7.8, 4.2 Hz, 1H), 4.01–3.87

(m, 2H), 3.73 (s, 6H), 2.39–2.18 (m, 2H), 1.70–1.52 (m,

2H), 1.37–1.24 (m, 8H), 0.92–0.85 (m, 3H); MS (FAB,

Pos.) m/z 448 (M+H)⁺, 427, 426, 405; HRMS

(MALDI-TOF, Pos.) calcd for free acid form

C₁₈H₃₀NO₇P+Na⁺: 426.1658; found: 426.1705.

2 - (3 - Methoxyphenyl) - 2 - octanoylaminoethyl disodium

phosphate (23b). White powder; TLC R_f=0.28 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3278, 2955, 2930, 2858,

1632, 1551, 1491, 1466, 1436, 1377, 1262, 1103 cm^ꢀ1; ¹H

NMR (300 MHz, CD₃OD) d 7.18 (dd, J=8.1, 8.1 Hz,

1H), 6.95–6.88 (m, 2H), 6.78–6.72 (m, 1H), 4.95–4.80

(m, 1H), 4.02–3.85 (m, 2H), 3.77 (s, 3H), 2.40–2.20 (m,

2H), 1.70–1.50 (m, 2H), 1.40–1.20 (m, 8H), 0.89 (t,

J=6.3 Hz, 3H); MS (FAB, Pos.) m/z 440 (M+Na)⁺,

Preparation of 25a–c and 47: the following compounds

were prepared according to the same procedures as

described for the preparation of 22 from 60t (general

method C).

418 (M+H)⁺, 396; HRMS (MALDI-TOF, Pos.) calcd

+

.

for C₁₇H₂₆NO₆P 2Na+H : 418.1371; found: 418.1355.

2 - (4 - Methoxyphenyl) - 2 - octanoylaminoethyl disodium

phosphate (23c). White powder; TLC R_f=0.28 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3285, 2930, 2858, 1631,

1550, 1515, 1465, 1250, 1102 cm^ꢀ1; ¹H NMR (300 MHz,

CD₃OD) d 7.26 (d, J=8.7 Hz, 2H), 6.82 (d, J=8.7Hz,

2H), 4.92–4.80 (m, 1H), 4.00–3.85 (m, 1H), 3.75 (s, 3H),

2.38–2.18 (m, 2H), 1.68–1.50 (m, 2H), 1.40–1.20 (m,

8H), 0.89 (t, J=6.5 Hz, 3H); MS (FAB, Pos.) m/z 440

2-(2-Chlorophenyl)-2-octanoylaminoethyl disodium phos-

phate (25a). Ivorypowder; TLC R_f=0.26 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3284, 3066, 2955, 2928,

2857, 2359, 1651, 1541, 1467, 1442, 1202, 1058 cm^ꢀ1; ¹H

NMR (200 MHz, CD₃OD) d 7.46–7.42 (m, 1H), 7.38–

7.33 (m, 1H), 7.29–7.20 (m, 2H), 5.44 (dd, J=7.0 Hz,

4.0 Hz, 1H), 4.15–3.90 (m, 2H), 2.32–2.24 (m, 2H),

1.65–1.54 (m, 2H), 1.36–1.22 (m, 8H), 0.88 (t, J=7.0

Hz, 3H); MS (FAB, Pos.) m/z 422 (M+H)⁺; HRMS

(MALDI-TOF, Pos.) calcd for C₁₆H₂₃ClNO₅P^ꢃ

2Na+H⁺: 422.0876; found: 422.0924.

(M+Na)⁺, 418 (M+H)⁺, 396; HRMS (MALDI-TOF,

+

.

Pos.) calcd for C₁₇H₂₆NO₆P 2Na+H : 418.1371;

found: 418.1323.

2-(2-Methylphenyl)-2-octanoylaminoethyl disodium phos-

phate (24a). White powder; TLC R_f=0.33 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3272, 2928, 1623, 1557,

2-(3-Chlorophenyl)-2-octanoylaminoethyl disodium phos-

phate (25b). Oﬀ-white powder; TLC R_f=0.33 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3429, 1645, 1550, 1465,

1

1095, 988 cm^ꢀ1; H NMR (300 MHz, CD₃OD) d 7.33–

1204, 1058 cm^ꢀ1; H NMR (300 MHz, CD₃OD) d 7.37

7.28 (m, 1H), 7.13–7.05 (m, 3H), 5.12 (t, J=6.3 Hz, 1H),

3.87 (t like, J=6.3 Hz, 1H), 2.43 (s, 3H), 2.36–2.17 (m,

2H), 1.64–1.52 (m, 2H), 1.33–1.22 (m, 8H), 0.88 (brt,

J=6.6 Hz, 3H); MS (FAB, Pos.) m/z 402 (M+H)⁺, 380,

358; HRMS (MALDI-TOF, Pos.) calcd for free acid

form C₁₇H₂₈NO₅P+Na⁺: 380.1603; found: 380.1611.

(s, 1H), 7.30–7.20 (m, 3H), 5.02 (dd, J=6.9, 4.2 Hz,

1H), 4.10–3.90 (m, 2H), 2.35–2.13 (m, 2H), 1.70–1.55

(m, 2H), 1.40–1.10 (m, 8H), 0.88 (t, J=6.6 Hz, 3H); MS

(FAB. Pos.) m/z 422 (M+H)⁺, 400; HRMS (MALDI-

+

.

TOF, Pos.) calcd for C₁₆H₂₃ClNO₅P 2Na+H :

422.0876; found: 422.0899.

2-(3-Methylphenyl)-2-octanoylaminoethyl disodium phos-

phate (24b). Oﬀ-white powder; TLC R_f=0.33 (CHCl₃/

2-(4-Chlorophenyl)-2-octanoylaminoethyl disodium phos-

phate (25c). White powder; TLC R_f=0.39 (CHCl₃/

3774

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

MeOH/H₂O, 65/35/8); IR (KBr) 3285, 2928, 2857, 1647,

1549, 1494, 1466, 1202, 1092, 1056, 1015 cm^ꢀ^{1 1}H

NMR (300 MHz, CD₃OD) d 6.86 (d, J=1.4 Hz, 1H),

6.81 (dd, J=8.2, 1.4 Hz, 1H), 6.70 (d, J=8.2 Hz, 1H),

5.87 (s, 2H), 4.80 (dd, J=7.6, 4.8 Hz, 1H), 4.00–3.81 (m,

2H), 2.38–2.14 (m, 2H), 1.70–1.50 (m, 2H), 1.35–1.20

(m, 8H), 0.95–0.83 (m, 3H); MS (FAB, Pos.) m/z 432

(M+H)⁺, HRMS (MALDI-TOF, Pos.) calcd for

C₁₇H₂₄NO₇P^ꢃ2Na+H⁺: 432.1164; found; 432.1143.

;

NMR (300 MHz, CD₃OD) d 7.36–7.33 (m, 2H), 7.31–

7.27 (m, 2H), 5.05–5.01 (m, 1H), 4.08–3.93 (m, 2H),

2.28–2.23 (m, 2H), 1.64–1.57 (m, 2H), 1.30–1.27 (m,

8H), 0.91–0.86 (m, 3H); MS (FAB, Pos.) m/z 422

(M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd for

+

.

C₁₆H₂₃ClNO₅P 2Na+H : 422.0876; found: 422.0905.

2-Octanoylamino-2-(thiophen-2-yl)ethyl disodium phos-

phate (57). Beige amorphous powder; TLC R_f=0.33

(CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3276, 1634,

2-(3-Methylthiophenyl)-2-octanoylaminoethyl disodium

phosphate (47). White powder; TLC R_f=0.37 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3261, 1625, 1553, 1093,

1

1548, 1103, 989 cm^ꢀ1; H NMR (200 MHz, CD₃OD) d

988 cm^ꢀ1

;

¹H NMR (200 MHz, CD₃OD) d 7.25 (m,

7.20 (dd, J=5.0, 1.4 Hz, 1H), 7.03 (brd, J=3.4 Hz, 1H),

6.91 (dd, J=5.0, 3.4 Hz, 1H), 5.22 (dd, J=7.4, 4.8 Hz,

1H), 4.15–3.97 (m, 2H), 2.38–2.13 (m, 2H), 1.70–1.50

(m, 2H), 1.40–1.20 (m, 8H), 0.93–0.82 (m, 3H); MS

(FAB, Pos.) m/z 394 (M+H)⁺,416, 372; HRMS

1H), 7.18 (d, J=6.8 Hz, 1H), 7.14–7.07 (m, 2H), 4.89–

4.83 (m, 1H), 4.04–3.85 (m, 2H), 2.42–2.16 (m, 2H), 2.45

(s, 3H), 1.68–1.52 (m, 2H), 1.35–1.24 (m, 8H), 0.92–0.84

(m, 3H); MS (FAB, Pos.) m/z 434 (M+H)⁺, 456, 412;

HRMS (MALDI-TOF, Pos.) calcd for C₁₇H₂₆NO₅

.

(MALDI-TOF, Pos.) calcd for C₁₄H₂₂NO₅PS

2Na+H⁺: 394.0830; found; 394.0801.

+

.

PS 2Na+H : 434.1143; found: 434.1154.

Preparation of 53–57: the following compounds were

prepared according to the same procedures as described

for the preparation of 1 from 60e (general method A).

Methyl (3-hydroxyphenyl)(octanoylamino)acetate (70).

Methyl amino(3-hydroxyphenyl)acetate was prepared

from 3-benzyloxy benzaldehyde according to the conven-

tional procedures.¹⁶To a stirred mixture of methyl

amino(3-hydroxyphenyl)acetate (15.7 g, 86.7 mmol) and

NaHCO₃(21.8 g, 260 mmol) in THF (170 mL) was added

dropwise n-octanoyl chloride (16.3 mL, 95.3 mmol) at

0 ^ꢁC. Stirring was continued for 30 min at that tempera-

ture. The reaction mixture was poured into ice-water and

extracted with EtOAc/THF. The organic layer was washed

with brine and dried over MgSO₄. Removal of the solvent

byevaporation aﬀorded a crude product, which was

washed with Et₂O/n-hexane to give 70: beige powder; 88%

2-(Naphtalen-1-yl)-2-octanoylaminoethyl disodium phos-

phate (53). White powder; TLC R_f=0.33 (CHCl₃/

MeOH/H₂O, 65/25/8); IR (KBr) 3418, 3051, 2955, 2930,

1645, 1540, 1465, 1212, 1057 cm^ꢀ1; ¹H NMR (300 MHz,

CD₃OD) d 8.21 (d, J=8.4 Hz, 1H), 7.88–7.76 (m, 2H),

7.59–7.41 (m, 4H), 5.95 (dd, J=7.5 Hz, 4.5 Hz, 1H),

4.31–4.23 (m, 1H), 4.14–4.06 (m, 1H), 2.37–2.22 (m,

2H), 1.68–1.55 (m, 2H), 1.29–1.26 (m, 8H), 0.89–0.85

(m, 3H); MS (FAB. Pos.) m/z 438 (M+H)⁺; HRMS

+

1

.

(MALDI-TOF, Pos.) calcd for C₂₀H₂₆NO₅P 2Na+H :

438.1422; found: 438.1412.

yield; TLC R_f=0.29 (n-hexane/EtOAc, 2/1); H NMR

(300 MHz, CDCl₃) d 7.19 (t, J=7.8 Hz, 1H), 6.91 (brs,

1H), 6.83–6.77 (m, 2H), 6.63 (brd, J=7.5 Hz, 1H), 5.53 (d,

J=7.5 Hz, 1H), 3.73 (s, 3H), 2.26 (t, J=8.1 Hz, 2H), 1.70–

1.55 (m, 2H), 1.31–1.23 (m, 8H), 0.88–0.80 (m, 3H).

2-(Naphtalen-2-yl)-2-octanoylaminoethyl disodium phos-

phate (54). White powder; TLC R_f=0.28 (CHCl₃/

MeOH/H₂O, 65/25/8); IR (KBr) 3851, 3428, 2954, 1626,

1555, 1464, 1300, 1105 cm^ꢀ1

;

¹H NMR (300MHz,

N-[2-Hydroxy-1-(3-methoxymethoxyphenyl)ethyl]octana-

mide (71a). To a stirred solution of 70 (922 mg, 3.0

mmol) and chloromethyl methyl ether (362 mg, 4.5

mmol) in THF (10 mL) was added NaH (60% disper-

sion in mineral oil, 132 mg, 3.3 mmol) at 0 ^ꢁC and stir-

ring was continued for 1 h at room temperature. The

reaction mixture was poured into satd NH₄Cl aq and

extracted with EtOAc. The organic layer was washed

with brine and dried over MgSO₄. Removal of the sol-

vent by evaporation gave methyl (3-methoxymethoxy)

phenyl(octanoylamino)acetate as a pale yellow powder,

to a stirred solution of which in THF (10 mL) was

added LiBH₄(131 mg, 6.0 mmol) at 0 ^ꢁC. Stirring was

continued for 18 h at room temperature. The reaction

mixture was quenched bysaturated NH ₄Cl aq and

extracted with EtOAc. The organic layer was washed

with brine and dried over MgSO_4.Removal of the sol-

vent byevaporation gave a residue, which was puriﬁed

bycolumn chromatographyon silica gel (Merck 7734,

n-hexane/EtOAc, 1/1) to aﬀord 71a: colorless oil; 80%

yield; TLC R_f=0.11 (n-hexane/EtOAc, 1/1).

CD₃OD) d 7.80–7.75 (m, 4H), 7.50 (dd, J=6.9 Hz, 1.5 Hz,

1H), 7.45–7.37 (m, 2H), 5.06 (dd, J=7.2 Hz, 3.6 Hz, 1H),

4.15–3.98 (m, 2H), 2.43–2.23 (m, 2H), 1.68–1.57 (m, 2H),

1.31–1.26 (m, 8H), 0.88–0.84 (m, 3H); MS (FAB. Pos.) m/

z 438 (M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd for

+

.

C₂₀H₂₆NO₅P 2Na+H : 438.1422; found; 438.1378.

2-(1,3-Dioxaindan-4-yl)-2-octanoylaminoethyl disodium

phosphate (55). Pale bitter orange powder; TLC

R_f=0.31 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3284,

;

2360, 1635, 1551, 1459, 1250, 1108 cm^ꢀ^{1 1}H NMR

(300 MHz, CD₃OD) d 6.81 (dd, J=7.5, 1.5 Hz, 1H),

6.73 (t, J=7.5 Hz, 1H), 6.66 (dd, J=7.5, 1.5 Hz, 1H),

5.93 (dd, J=3.9, 1.2 Hz, 2H), 5.03 (t, J=6.0 Hz, 1H),

4.01 (t, J=6.0 Hz, 2H), 2.29 (m, 2H), 1.59 (m, 2H), 1.40–

1.20 (m, 8H), 0.89 (t, J=6.6 Hz, 3H); MS (FAB. Pos.) m/

z 454 (M+Na)⁺; HRMS (MALDI-TOF, Pos.) calcd for

+

.

C₁₇H₂₄NO₇P 2Na+H : 432.1164; found; 432.1137.

2-(1,3-Dioxaindan-5-yl)-2-octanoylaminoethyl disodium

phosphate (56). Beige amorphous powder; TLC

R_f=0.35 (CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3285,

;

1631, 1550, 1505, 1491, 1249, 1101, 1044 cm^ꢀ^{1 1}H

N-[2-Hydroxy-1-(3-ethoxyphenyl)ethyl]octanamide (71b).

To a stirred solution of 70 (1.23 g, 4 mmol) in acetone

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3775

(10 mL) were added K₂CO₃(1.38 g, 10 mmol) and EtI

(1.25 g, 8 mmol) at room temperature and stirring was

continued for 8 h at the reﬂux temperature. After cool-

ing, removal of the precipitates byﬁltration through a

pad of Celite followed byevaporation of the ﬁltrate

gave an oilyresidue, which was dissolved in EtOAc. The

organic layer was successively washed with H₂O and

brine before being dried over MgSO₄which was con-

centrated to aﬀorded methyl (3-ethoxyphenyl)(octanoyl-

amino)acetate (1.3 g, 96%). The product was used for

the next reaction without further puriﬁcation: TLC

2H), 2.24 (brt, J=8.1 Hz, 2H), 2.17–1.96 (m, 1H),

1.70–1.50 (m, 2H), 1.40–1.20 (m, 8H), 0.87 (brt, J=6.6

Hz, 3H).

N-{2-Hydroxy-1-[3-(pentyloxy)phenyl]ethyl}octanamide

(71h). Yield, 69%; TLC R_f=0.24 (n-hexane/EtOAc, 1/

1

1); H NMR (300 MHz, CDCl₃) d 7.26 (dd, J=8.1, 8.1

Hz, 1H), 6.86–6.81 (m, 3H), 6.11 (brd, J=6.6 Hz, 1H),

5.02 (m, 1H), 3.94 (t, J=6.6 Hz, 2H), 3.92–3.80 (m, 2H),

2.77 (brs, 1H), 2.24 (brt, J=7.5 Hz, 2H), 1.82–1.60 (m,

4H), 1.50–1.20 (m, 12H), 0.93 (t, J=7.2 Hz, 3H), 0.87

(brt, J=6.9 Hz, 3H).

1

R_f=0.46 (n-hexane/EtOAc, 1/1); H NMR (300 MHz,

CDCl₃) d 7.25 (dd, J=8.4, 8.4 Hz, 1H), 6.92–6.82 (m,

3H), 6.38 (d, J=7.2 Hz, 1H), 5.55 (d, J=7.2 Hz, 1H),

4.02 (q, J=7.2 Hz, 3H), 3.72 (s, 3H), 2.23 (brt, J=8.4

Hz, 2H), 1.70–1.55 (m, 2H), 1.40 (t, J=7.2 Hz, 3H),

1.35–1.20 (m, 8H), 0.86 (brt, J=7.2 Hz, 3H). Com-

pound 71b was prepared according to the same proce-

dure as described for the preparation of 71a from 70

and puriﬁed bycolumn chromatographyon silica gel

(FL60D, n-hexane/EtOAc, 1/1–2/3): 74% yield; TLC

N-{1-[3-(Hexyloxy)phenyl]-2-hydroxyethyl}octanamide

(71i). Yield 87%; TLC R_f=0.28 (n-hexane/EtOAc, 1/

1); H NMR (300 MHz, CDCl₃) d 7.26 (dd, J=8.1, 8.1

Hz, 1H), 6.86–6.81 (m, 3H), 6.10 (brd, J=7.2 Hz, 1H),

5.03 (m, 1H), 3.94 (t, J=6.6 Hz, 2H), 3.92–3.80 (m, 2H),

2.77 (brs, 1H), 2.24 (brt, J=7.5 Hz, 2H), 1.82–1.60 (m,

4H), 1.50–1.20 (m, 12H), 0.91 (t, J=6.9 Hz, 3H), 0.87

(brt, J=7.2 Hz, 3H).

1

R_f=0.10 (n-hexane/EtOAc, 1/1); H NMR (300 MHz,

CDCl₃) d 7.26 (dd, J=8.4, 8.4 Hz, 1H), 6.86–6.80 (m,

3H), 6.11 (brd, J=6.9 Hz, 1H), 5.02 (m, 1H), 4.02 (q,

J=7.2 Hz, 3H), 3.91–3.81 (m, 2H), 2.23 (brt, J=7.5 Hz,

2H), 1.70–1.60 (m, 2H), 1.40 (t, J=7.2 Hz, 3H), 1.30–

1.20 (m, 8H), 0.87 (brt, J=6.9 Hz, 3H).

N-{1-[3-(Cyclopentyloxy)phenyl]-2-hydroxyethyl}octan-

amide (71k). Yield, 61%; TLC R_f=0.42 (n-hexane/

EtOAc, 1/1); H NMR (300 MHz, CDCl₃) d 7.25 (dd,

J=7.5, 7.5 Hz, 1H), 6.84–6.78 (m, 3H), 6.11 (brd,

J=6.9 Hz, 1H), 5.01 (m, 1H), 4.74 (m, 1H), 3.92–3.80

(m, 2H), 2.83 (brs, 1H), 2.24 (brt, J=8.1 Hz, 2H), 2.00–

1.55 (m, 10H), 1.40–1.20 (m, 8H), 0.87 (brt, J=6.9 Hz,

3H).

1

Preparation of 71c–71i and 71k–71m: the following

compounds were prepared according to the same pro-

cedures as described for the preparation of 71b from 70.

N - (1 - {3- [2 - (Dimethylamino) - 2 - oxoethoxy]phenyl} - 2-

hydroxyethyl) octanamide (71l). Yield, 82%; TLC

R_f=0.14 (EtOAc/MeOH, 19/1); ¹H NMR (300 MHz,

CDCl₃) d 7.25 (t, J=8.1 Hz, 1H), 6.92–6.81 (m, 3H),

6.32 (d, J=6.9 Hz, 1H), 5.04–4.99 (m, 1H), 4.67 (s, 2H),

3.85–383 (m, 2H), 3.07 (s. 3H), 2.96 (s, 3H), 2.23 (t,

J=7.2 Hz, 2H), 1.68–1.58 (m, 2H), 1.29–1.23 (m, 8H),

0.89–0.84 (m, 3H).

N-[2-Hydroxy-1-(3-propyloxyphenyl)ethyl]octanamide

(71c). Yield, 66%; TLC R_f=0.40 (n-hexane/EtOAc, 1/

1); H NMR (300 MHz, CDCl₃) d 7.26 (dd, J=7.5, 7.5

Hz, 1H), 6.85–6.80 (m, 3H), 6.12 (brd, J=6.9 Hz, 1H),

5.02 (m, 1H), 3.90 (t, J=7.2 Hz, 2H), 3.85 (m, 2H), 2.23

(brt, J=7.8 Hz, 2H), 1.85–1.70 (m, 2H), 1.70–1.57 (m,

2H), 1.40–1.20 (m, 8H), 1.03 (t, J=7.8 Hz, 3H), 0.87

(brt, J=6.9 Hz, 3H).

1

N-{1-[3-(3-Benzyloxy)phenyl]-2-hydroxyethyl}octanamide

(71m). Yield, 79%; white powder; TLC R_f=0.25

(n-hexane/toluene, 3/1); H NMR (300 MHz, CDCl₃) d

7.44–7.25 (m, 6H), 6.92–6.87 (m, 3H), 6.07 (d, J=6.9

Hz, 1H), 5.05 (s, 2H), 5.02 (t, J=9.0 Hz, 1H), 3.92–3.80

(m, 2H), 2.23 (t, J=7.8 Hz, 2H), 1.69–1.59 (m, 2H),

1.30–1.26 (m, 8H), 0.86 (t, J=6.9 Hz, 3H).

N-[2-Hydroxy-1-(3-isopropyloxyphenyl)ethyl]octanamide

(71d). Yield, 68%; TLC R_f=0.40 (n-hexane/EtOAc, 1/

1); H NMR (300 MHz, CDCl₃) d 7.25 (dd, J=7.5, 7.5

Hz, 1H), 6.84–6.79 (m, 3H), 6.13 (brd, J=6.6 Hz, 1H),

5.00 (m, 1H), 4.53 (m, 1H), 3.84 (m, 2H), 2.23 (brt,

J=8.1 Hz, 2H), 1.70–1.60 (m, 2H), 1.32 (d, J=6.3 Hz,

6H), 1.32–1.22 (m, 8H), 0.86(t, J=6.9 Hz, 3H).

1

N-{1-[3-(Cyclobutyloxy)phenyl]-2-hydroxyethyl}octan-

amide (71j). To a stirred mixture of 70 (614 mg, 2

mmol), PPh₃(524 mg, 2 mmol) and cyclobutyl alcohol

(144 mg, 2 mmol) in THF (6 mL) was added dropwise

DEAD (0.32 mL, 2 mmol) at 0 ^ꢁC. Stirring was con-

tinued for 24 h at room temperature. Removal of the

solvent byevaporation gave a residue, which was pur-

iﬁed bycolumn chromatographyon silica gel (Merck

7734, n-hexane/EtOAc, 4/1–3/1) to aﬀorded methyl

[(3-cyclobutyloxyphenyl)](octanoylamino)acetate (400

mg, 55%): TLC R_f=0.33 (n-hexane/EtOAc, 3/1); ¹H

NMR (300 MHz, CD₃OD) d 7.23 (dd, J=7.8, 7.8 Hz,

1H), 6.90 (brd, J=7.8 Hz, 1H), 6.80 (brt, J=2.1 Hz,

1H), 6.75 (dd, J=7.8, 2.1 Hz, 1H), 6.35 (brd, J=7.5 Hz,

1H), 5.54 (d, J=7.2 Hz, 1H), 4.62 (m, 1H), 3.72 (s, 3H),

N-{1-[3-(Butyloxy)phenyl]-2-hydroxyethyl}octanamide

(71e). Yield, 75%; TLC R_f=0.24 (n-hexane/EtOAc, 1/

1); H NMR (300 MHz, CDCl₃) d 7.26 (dd, J=7.8, 7.8

Hz, 1H), 6.86–6.81 (m, 3H), 6.10 (brd, J=6.6 Hz, 1H),

5.03 (m, 1H), 3.95 (t, J=6.3 Hz, 2H), 3.92–3.80 (m, 2H),

2.76 (brs, 1H), 2.24 (brt, J=7.5 Hz, 2H), 1.80–1.40 (m,

6H), 1.40–1.20 (m, 8H), 0.98 (t, J=7.5 Hz, 3H), 0.87

(brt, J=6.9 Hz, 3H).

1

N-{2-Hydroxy-1-[3-(isobutyloxy)phenyl]ethyl}octanamide

(71f). Yield, 59%; TLC R_f=0.40 (n-hexane/EtOAc, 1/

1

1); H NMR (300 MHz, CDCl₃) d 7.25 (dd, J=7.2, 7.2

Hz, 1H), 6.86–6.81 (m, 3H), 6.11 (brd, J=6.6 Hz, 1H),

5.02 (m, 1H), 3.92–3.80 (m, 2H), 3.70 (d, J=6.3 Hz,

3776

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

2.50–2.37 (m, 2H), 2.25–2.10 (m, 4H), 1.90–1.55 (m,

4H), 1.35–1.20 (m, 8H), 0.87 (t, J=6.9 Hz, 3H). The

compound 71j was prepared from methyl (3-

cyclobutyloxyphenyl)(octanoylamino)acetate according

to the same procedures as described for the preparation of

71a from methyl (3-methoxymethoxy)phenyl(octanoyl-

amino) acetate: TLC R_f=0.36 (n-hexane/EtOAc, 1/1);

¹H NMR (300 MHz, CDCl₃) d 7.24 (dd, J=7.8, 7.8 Hz,

1H), 6.84 (brd, J=7.8 Hz, 1H), 6.75–6.70 (m, 2H), 6.10

(brd, J=6.6 Hz, 1H), 5.01 (m, 1H), 4.62 (m, 1H), 3.91–

3.82 (m, 2H), 2.50–2.35 (m, 2H), 2.25–2.10 (m, 4H),

1.90–1.60 (m, 4H), 1.40–1.20 (m, 8H), 0.87 (t, J=6.9

Hz, 3H).

2-octanoylaminoethyl dihydrogen phosphate was

obtained from 70a according to the same procedures as

described for the preparation of 27 from 70a, which was

converted to the disodium salt according to the same

procedures as described for the preparation of 1 from

62e: oﬀ-white amorphous powder; TLC R_f=0.28

(CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3407, 1628,

1

1559, 1099, 986 cm^ꢀ1; H NMR; (300 MHz, CD₃OD) d

7.18 (t, J=7.8 Hz, 1H), 7.01 (brs, 1H), 6.98 (brd, J=7.8

Hz, 1H), 6.86 (ddd, J=7.8, 2.1, 0.9 Hz, 1H), 5.16 (d,

J=6.6 Hz, 1H), 5.13 (d, J=6.6 Hz, 1H), 4.88–4.82 (m,

1H), 4.01–3.87 (m, 2H), 3.42 (s, 3H), 2.38–2.18 (m, 2H),

1.69–1.53 (m, 2H), 1.36–1.24 (m, 8H), 0.91–0.86 (m,

3H); MS (FAB, Pos.) m/z 448 (M+H)⁺, 426, 404;

.

N-{1-[3-(1-Ethylpropoxy)phenyl]-2-hydroxyethyl}octan-

amide (71g). The title compound 71g was prepared

from 70 according to the same procedures as described for

the preparation of 71j from 70 and puriﬁed bycolumn

chromatographyon silica gel (Merck 7734, n-hexane/

EtOAc, 4/1–3/1) to give methyl [3-(3-pentyloxyl-

phenyl)](octanoylamino)acetate: 48% yield; TLC

HRMS (MALDI-TOF, Pos.) calcd for C₁₈H₂₈NO₇P

2Na+H⁺: 448.1477; found: 448.1488.

Preparation of 28–37: the following compounds were

prepared according to the same procedure as described

for the preparation of 1 from 60e (general method A).

1

R_f=0.38 (n-hexane/EtOAc, 3/1); H NMR (300 MHz,

2-(3-Ethoxyphenyl)-2-octanoylaminoethyl disodium phos-

phate (28). Yield, 36%; colorless powder; TLC

R_f=0.22 (CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3286,

2928, 1637, 1547, 1261, 1116 cm^ꢀ1; ¹H NMR (300 MHz,

CD₃OD) d 7.15 (t, J=8.1 Hz, 1H), 6.89 (m, 2H), 6.72

(dd, J=8.1, 1.8 Hz, 1H), 4.83 (m, 1H), 4.05–3.87 (m,

4H), 2.38–2.18 (m, 2H), 1.61 (m, 2H), 1.35 (t, J=7.2 Hz,

3H), 1.30–1.20 (m, 8H), 0.88 (brt, J=6.6 Hz, 3H); MS

(FAB, Pos.) m/z 432 (M+H)⁺; HRMS (MALDI-TOF,

Pos.) calcd for C₁₈H₂₈NO₆P^ꢃ2Na+H⁺: 432.1528;

found: 432.1518.

CDCl₃) d 7.23 (dd, J=8.1, 8.1 Hz, 1H), 6.89–6.81 (m,

3H), 6.33 (brd, J=7.2 Hz, 1H), 5.55 (d, J=7.2 Hz, 1H),

4.10 (m, 1H), 3.73 (s, 3H), 2.23 (t, J=8.4 Hz, 2H), 1.70–

1.57 (m, 6H), 1.35–1.20 (m, 8H), 0.94 (t, J=7.2 Hz, 6H),

0.86 (brt, J=6.9 Hz, 3H). Compound 71g: 98% yield;

TLC R_f=0.45 (n-hexane/EtOAc, 1/1); ¹H NMR

(300 MHz, CDCl₃) d 7.25 (dd, J=9.0, 7.5 Hz, 1H),

6.84–6.79 (m, 3H), 6.09 (brd, J=6.6 Hz, 1H), 5.03 (m,

1H), 4.11 (m, 1H), 3.90–3.83 (m, 2H), 2.24 (t, J=7.5 Hz,

2H), 1.70–1.60 (m, 6H), 1.40–1.20 (m, 8H), 0.95 (t,

J=7.5 Hz, 6H), 0.87 (brt, J=6.9 Hz, 3H).

2-Octanoylamino-2-(3-propyloxyphenyl)ethyl disodium

phosphate (29). Colorless powder; TLC R_f=0.22

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3278, 2928,

1632, 1550, 1452, 1264, 1111 cm^ꢀ1; ¹H NMR (300 MHz,

CD₃OD) d 7.15 (t, J=8.1 Hz, 1H), 6.89 (m, 2H), 6.72

(dd, J=8.1, 1.8 Hz, 1H), 4.83 (m, 1H), 4.00–3.87 (m,

4H), 2.38–2.18 (m, 2H), 1.76 (tq, J=7.5, 7.5 Hz, 2H),

1.61 (m, 2H), 1.38–1.20 (m, 8H), 1.02 (t, J=7.5 Hz, 3H),

0.88 (brt, J=6.6 Hz, 3H); MS (FAB, Pos.) m/z 446

2-(3-Hydoxyphenyl)-2-octanoylaminoethyl disodium phos-

phate (27). 2-(3-Methoxymethoxyphenyl)-2-octanoyl-

aminoethyl dihydrogen phosphate was prepared from

71a according to essentiallythe same procedures as

described for the preparation of 62e from 60e. To a

solution of 2-(3-metoxymetylphenyl)-2-octanoylamino-

ethyl dihydrogen phosphate in MeOH (10 mL) was

added a few drops of 6 M HCl and the mixture was

stirred at room temperature for 18 h. Following the

addition of H₂O, the mixture was extracted with

EtOAc. The organic layer was washed with brine and

dried over Na₂SO₄. Removal of the solvent byeva-

poration gave 2-(3-hydroxyphenyl)-2-octanoylamino-

ethyl dihydrogen phosphate as a white amorphous

powder (54% yield), which was converted to the dis-

odium salt according to the same procedures as descri-

bed for the preparation of 1 from 62e: grayamorphous

powder; TLC R_f=0.22 (CHCl₃/MeOH/H₂O, 65/35/8);

(M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd for

+

.

C₁₉H₃₀NO₆P 2Na+H : 446.1684; found: 446.1719.

2-(3-Isopropyloxyphenyl)-2-octanoylaminoethyl disodium

phosphate (30). Colorless powder; TLC R_f=0.22

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3281, 2928,

1634, 1550, 1489, 1259, 1117 cm^ꢀ1; ¹H NMR (300 MHz,

CD₃OD) d 7.14 (t, J=8.1 Hz, 1H), 6.89 (m, 2H), 6.72

(dd, J=8.1, 2.4 Hz, 1H), 4.83 (m, 1H), 4.56 (qq, J=6.0,

6.0 Hz, 1H), 4.00–3.87 (m, 2H), 2.38–2.18 (m, 2H), 1.62

(m, 2H), 1.38–1.20 (m, 8H), 1.28 (d, J=6.0 Hz, 3H),

1.26 (d, J=6.0 Hz, 3H), 0.84 (brt, J=6.6 Hz, 3H); MS

1

IR (KBr) 3398, 1621, 1544, 1460, 1089, 983 cm^ꢀ1; H

NMR (300 MHz, CD₃OD) d 7.04 (t, J=7.8 Hz, 1H),

6.78 (brs, 1H), 6.74 (brd, J=7.8 Hz, 1H), 6.59 (dd, J=7.8,

2.4 Hz, 1H), 4.82 (dd, J=8.1, 3.9 Hz, 1H), 4.00–3.86 (m,

2H), 2.36–2.18 (m, 2H), 1.68–1.53 (m, 2H), 1.35–1.24 (m,

8H), 0.91–0.86 (m, 3H); MS (FAB, Pos.) m/z 404

(FAB, Pos.) m/z 446 (M+H)⁺; HRMS (MALDI-TOF,

+

.

Pos.) calcd for C₁₉H₃₀NO₆P 2Na+H : 446.1684;

found: 446.1720.

(M+H)⁺, 426, 382; HRMS (MALDI-TOF, Pos.) calcd

2-(3-Butyloxyphenyl)-2-octanoylaminoethyl disodium

phosphate (31). Colorless powder; TLC R_f=0.35

+

.

for C₁₆H₂₄NO₆P 2Na+H : 404.1215; found: 404.1255.

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3260, 2930,

1658, 1626, 1555, 1460, 1097, 990 cm^ꢀ^{1 1}H NMR

;

(300 MHz, CD₃OD) d 7.15 (t, J=8.1 Hz, 1H), 6.89 (m,

2-(3-Methoxymethoxyphenyl)-2-octanoylaminoethyl dis-

odium phosphate (40). 2-[(3-Methoxymethoxyphenyl)]-

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3777

2H), 6.72 (dd, J=8.1, 2.4 Hz, 1H), 4.83 (m, 1H),

4.00–3.87 (m, 4H), 2.40–2.18 (m, 2H), 1.77–1.40 (m,

6H), 1.40–1.20 (m, 8H), 0.97 (t, J=7.5 Hz, 3H), 0.88

(brt, J=6.3 Hz, 3H); MS (FAB, Pos.) m/z 482

1H), 6.79 (brt, J=1.8 Hz, 1H), 6.63 (dd, J=7.8, 1.8 Hz,

1H), 4.84 (m, 1H), 4.64 (m, 1H), 3.98–3.86 (m, 2H),

2.50–2.00 (m, 6H), 1.85–1.50 (m, 4H), 1.40–1.20 (m,

8H), 0.88 (brt, J=6.6 Hz, 3H); MS (FAB, Pos.) m/z 480

(M+Na)⁺, 460 (M+H)⁺; HRMS (MALDI-TOF,

(M+Na)⁺, 458 (M+H)⁺; HRMS (MALDI-TOF, Pos.)

+

.

Pos.) calcd for C₂₀H₃₂NO₆P 2Na+H : 460.1841;

found: 460.1837.

.

calcd for C₂₀H₃₀NO₆P 2Na+H : 458.1684; found:

458.1716.

2-(3-Isobutyloxyphenyl)-2-octanoylaminoethyl disodium

phosphate (32). Colorless powder; TLC R_f=0.26

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3298, 2928,

2-(3-Cyclopentyloxyphenyl)-2-octanoylaminoethyl diso-

dium phosphate (37). Colorless powder; TLC R_f=0.26

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3288, 2928,

1

1631, 1550, 1469, 1266, 1113, 988 cm^ꢀ1

;

¹H NMR

1636, 1550, 1450, 1112, 987 cm^ꢀ1; H NMR (300 MHz,

(300 MHz, CD₃OD) d 7.15 (t, J=7.8 Hz, 1H), 6.89 (m,

2H), 6.73 (dd, J=7.8, 1.8 Hz, 1H), 4.84 (m, 1H), 4.00–

3.88 (m, 2H), 3.75–3.65 (m, 2H), 2.39–2.18 (m, 2H),

2.09–1.95 (m, 1H), 1.70–1.50 (m, 2H), 1.40–1.20 (m,

8H), 1.01 (d, J=6.9 Hz, 6H), 0.88 (brt, J=6.6 Hz, 3H);

MS (FAB, Pos.) m/z 482 (M+Na)⁺, 460 (M+H)⁺;

CD₃OD) d 7.14 (t, J=7.8 Hz, 1H), 6.87 (m, 2H), 6.69

(dd, J=7.8, 1.8 Hz, 1H), 4.84 (m, 1H), 4.77 (m, 1H),

4.00–3.87 (m, 2H), 2.40–2.18 (m, 2H), 1.95–1.50 (m,

10H), 1.40–1.20 (m, 8H), 0.88 (brt, J=6.6 Hz, 3H); MS

(FAB, Pos.) m/z 494 (M+Na)⁺, 472 (M+H)⁺; HRMS

+

.

(MALDI-TOF, Pos.) calcd for C₂₁H₃₂NO₆P 2Na+H :

472.1841; found: 472.1838.

.

HRMS (MALDI-TOF, Pos.) calcd for C₂₀H₃₂NO₆P

2Na+H⁺: 460.1841; found: 460.1850.

2-(3-Dimethylaminocarbonylmethyloxyphenyl)-2-octanoyl-

aminoethyl disodium phosphate (38). Ivorypowder; TLC

R_f=0.30 (CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3855,

3424, 2928, 1648, 1544, 1491, 1458, 1363, 1257, 1090

2-[3-(3-Pentyloxyphenyl)]-2-octanoylaminoethyl disodium

phosphate (33). Colorless powder; TLC R_f=0.23

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3279, 2962,

1

2929, 1632, 1551, 1463, 1268, 1111 cm^ꢀ1

;

¹H NMR

cm^ꢀ1; H NMR (300 MHz, CD₃OD) d 7.18 (t, J=8.1

(300 MHz, CD₃OD) d 7.14 (t, J=8.1 Hz, 1H), 6.88 (m,

2H), 6.72 (dd, J=8.1, 1.8 Hz, 1H), 4.84 (m, 1H), 4.14

(m, 1H), 4.00–3.87 (m, 2H), 2.40–2.18 (m, 2H), 1.70–

1.50 (m, 6H), 1.40–1.20 (m, 8H), 0.94 (t, J=7.5 Hz, 3H),

0.93 (t, J=7.5 Hz, 3H), 0.88 (brt, J=6.9 Hz, 3H); MS

Hz, 1H), 6.98–6.96 (m, 2H), 6.83–6.80 (m, 1H), 4.88–

4.84 (m, 1H), 4.75 (s, 2H), 3.99–3.91 (m, 2H), 3.09 (s,

3H), 2.96 (s, 3H), 2.38–2.18 (m, 2H), 1.66–1.53 (m, 2H),

1.30–1.28 (m, 8H), 0.88 (t, J=6.9 Hz, 3H); MS (FAB.

Pos.) m/z 489 (M+H)⁺; HRMS (MALDI-TOF, Pos.)

+

(FAB, Pos.) m/z 496 (M+Na)⁺, 474 (M+H)⁺; HRMS

calcd for C₂₀H₃₁N₂O₇P 2Na+H : 489.1743; found:

489.1748.

.

+

.

(MALDI-TOF, Pos.) calcd for C₂₁H₃₄NO₆P 2Na+H :

474.1997; found: 474.1992.

Ethyl

{3-[2-hydroxy-1-(octanoylamino)ethyl]phenoxy}

2-(3-n-Pentyloxyphenyl)-2-octanoylaminoethyl disodium

phosphate (34). Colorless powder; TLC R_f=0.35

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3259, 2928,

acetate (72). A mixture of 71m (1.2 g, 3.25 mmol) in

EtOH (20 mL) and 10% Pd-C (120 mg) was stirred at

room temperature under an atmospheric pressure of

hydrogen for 2 h. Removal of the catalyst by ﬁltration

through a pad of Celite followed byevaporation of the

ﬁltrate aﬀorded 2-(3-hydroxyphenyl)-2-octanoylami-

noethanol as an oilyresidue which was used for the next

reaction without further puriﬁcation: TLC R_f=0.50

1656, 1625, 1556, 1460, 1098, 990 cm^ꢀ^{1 1}H NMR

;

(300 MHz, CD₃OD) d 7.15 (t, J=8.1 Hz, 1H), 6.89 (m,

2H), 6.72 (dd, J=8.1, 1.8 Hz, 1H), 4.83 (m, 1H), 4.00–

3.87 (m, 4H), 2.40–2.18 (m, 2H), 1.80–1.20 (m, 16H),

0.94 (t, J=7.2 Hz, 3H), 0.88 (brt, J=6.3 Hz, 3H); MS

(FAB, Pos.) m/z 496 (M+Na)⁺, 474 (M+H)⁺; HRMS

(EtOAc); H NMR (300 MHz, CDCl₃) d 7.25–7.18 (m,

1

+

.

(MALDI-TOF, Pos.) calcd for C₂₁H₃₄NO₆P 2Na+H :

474.1997; found: 474.2013.

1H), 6.83–6.75 (m, 3H), 6.20 (d, J=7.5 Hz, 1H), 4.98

(dt, J=7.5, 4.0 Hz, 1H), 3.85 (d, J=4.0 Hz, 2H), 2.25 (t,

J=7.5 Hz, 2H), 1.67 (m, 2H), 1.40–1.20 (m, 8H), 0.88 (t,

J=6.2 Hz, 3H). To a stirred mixture of 2-(3-hydroxy-

phenyl)-2-octanoylaminoethanol (906 mg, 3.25 mmol),

K₂CO₃(897 mg, 6.5 mmol) and KI (647 mg, 3.9 mmol)

in DMF (10 mL) was added ethyl bromoacetate (0.43

mL, 3.9 mmol) and stirring was continued at room

temperature for 2 days. The reaction mixture was

poured into ice-cooled 1 M HCl and extracted with

EtOAc. The organic layer was successively washed with

H₂O and brine before being dried over Na₂SO₄.

Removal of the solvent byevaporation gave a residue

which was puriﬁed bycolumn chromatographyon silica

gel (Merck 7734, n-hexane/EtOAc, 1/1–1/2–1/3) to

aﬀord 72 as a colorless wax: 81% yield in 2 steps; TLC

2-(3-n-Hexyloxyphenyl)-2-octanoylaminoethyl disodium

phosphate (35). Colorless powder; TLC R_f=0.35

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3259, 2927,

;

1657, 1625, 1556, 1459, 1099, 990 cm^ꢀ^{1 1}H NMR

(300 MHz, CD₃OD) d 7.15 (t, J=8.1 Hz, 1H), 6.89 (m,

2H), 6.73 (dd, J=8.1, 2.4 Hz, 1H), 4.83 (m, 1H), 4.00–

3.87 (m, 4H), 2.40–2.18 (m, 2H), 1.80–1.20 (m, 18H),

0.92 (t, J=6.9 Hz, 3H), 0.88 (brt, J=6.6 Hz, 3H); MS

(FAB, Pos.) m/z 488 (M+H)⁺; HRMS (MALDI-TOF,

+

.

Pos.) calcd for C₂₂H₃₆NO₆P 2Na+H : 488.2154;

found: 488.2179.

2-(3-Cyclobutyloxyphenyl)-2-octanoylaminoethyl disodium

phosphate (36). Colorless powder; TLC R_f=0.21

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3263, 2927,

1

R_f=0.55 (EtOAc); H NMR (300 MHz, CDCl₃) ꢀ 7.28

(t, J=8.0 Hz, 1H), 6.92 (d, J=8.0 Hz, 1H), 6.88 (t,

J=2.7 Hz, 1H), 6.80 (dd, J=8.0, 2.7 Hz, 1H), 6.21 (d,

J=6.6 Hz, 1H), 5.03 (dt, J=6.6, 5.0 Hz, 1H), 4.61 (s,

1

1626, 1553, 1453, 1101, 989 cm^ꢀ1; H NMR (300 MHz,

CD₃OD) d 7.13 (t, J=7.8 Hz, 1H), 6.89 (brd, J=7.8 Hz,

3778

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

2H), 4.27 (q, J=7.0 Hz, 2H), 3.85 (d, J=5.0 Hz, 2H),

2.24 (t, J=7.2 Hz, 2H), 1.64 (m, 2H), 1.40–1.20 (m) and

1.31 (t, J=7.0 Hz) total 11H, 0.88 (m, 3H).

into H₂O and extracted with EtOAc. The organic layer

was washed with brine and dried over MgSO₄. Removal

of the solvent by evaporation gave 2-(3-benzyloxy-

phenyl)-1-(tert-butyldimethylsilyl)-2-octanoylaminoethyl

ether as a colorless oil (15.2 g, quant.): TLC R_f=0.46

General method D

1

(n-hexane/EtOAc, 2/1); H NMR (200 MHz, CDCl₃) d

2-(3-Ethoxycarbonylmethylphenyl)-2-octanoylaminoethyl

disodium phosphate (39). To a stirred solution of 72 (960

mg, 2.6 mmol) and tetrazole (364 mg, 5.2 mmol) in

CH₃CN (10 mL) was added dibenzyl diisopropylpho-

sphoramidite (989 mg, 3.1 mmol) in CH₃CN (5 mL).

Stirring was continued at room temperature for 3 h. The

reaction mixture was poured into saturated NaHCO₃aq

and extracted with EtOAc. After the organic layer was

7.46–7.30 (m, 6H), 7.27–7.19 (m, 1H), 6.96–6.82 (m,

2H), 6.17 (d, J=7.8 Hz, 1H), 5.05 (s, 2H), 5.06–4.94 (m,

1H), 3.89 (dd, J=10.2, 4.4 Hz, 1H), 3.78 (dd, J=10.2,

4.4 Hz, 1H), 2.27–2.19 (m, 2H), 1.40–1.20 (m, H), 0.86

(s, 9H), 0.90–0.80 (m, 3H), ꢀ0.03 (s, 3H), ꢀ0.06 (s, 3H).

A

mixture of 2-(3-benzyloxyphenyl)-1-(tert-butyldi-

methylsilyl)-2-octanoylaminoethyl ether (15.23 g, 27.0

mmol) in MeOH (200 mL) and 10% Pd–C (1.0 g) was

stirred at room temperature under an atmospheric

pressure of hydrogen for 2 h. Removal of the catalyst by

ﬁltration through a pad of Celite followed byevapora-

tion aﬀorded 1-(tert-butyldimethylsilyl)-2-(3-hydroxy-

phenyl)-2-octanoylaminoethyl ether as a white powder

(10.6 g, quant.): TLC R_f=0.33 (n-hexane/EtOAc, 3/1);

¹H NMR (300 MHz, CDCl₃) d 7.11 (t, J=7.6 Hz, 1H),

6.80–6.66 (m, 3H), 6.35 (d, J=7.6 Hz, 1H), 4.96–4.88

(m, 1H), 3.87 (dd, J=10.2, 4.4 Hz, 1H), 3.72 (dd,

J=10.2, 4.4 Hz, 1H), 2.30–2.22 (m, 2H), 1.40–1.20 (m,

(H), 0.95–0.90 (m, 12H), ꢀ0.04 (s, 3H), ꢀ0.06 (s, 3H).

To a stirred solution of 1-(tert-butyldimethylsilyl)-2-(3-

hydroxyphenyl)-2-octanoylaminoethyl ether (8.26 g,

21.0 mmol) in pyridine (30 mL) was added dropwise

Tf₂O (3.90 mL, 23.1 mmol) at 0 ^ꢁC. Stirring was con-

tinued for 1 h at that temperature. The reaction mixture

was poured into 1 M HCl and extracted with EtOAc.

The organic layer was successively washed saturated

NaHCO₃aq and brine before being dried over MgSO₄.

Removal of the solvent byevaporation gave 1-( tert-

butyldimethylsilyl) - 2 - (3 - triﬂuoromethansulfonyloxy-

phenyl)-2-octanoylaminoethyl ether as a yellow oil (11.6 g,

quant.): TLC R_f=0.54 (n-hexane/EtOAc, 3/1). To a stir-

red mixture of 1-(tert-butyldimethylsilyl)-2-(3-triﬂuoro-

methansulfonyloxyphenyl)-2-octanoylaminoethyl ether

(9.46 g, 18 mmol), Et₃N (6.30 mL, 45.0 mmol) and

Pd(OAc)₂(202 mg, 0.9 mmol) in MeOH (36 mL)/

DMSO (54 mL) was added DPPP (371 mg, 0.9 mmol) at

room temperature. Stirring was continued at 70 ^ꢁC for 2

h under an atmospheric pressure of carbon monoxide.

The reaction mixture was poured into 0.2 M HCl and

extracted with EtOAc. The organic layer was succes-

successivelywashed with H O and brine, it was dried

2

over Na₂SO₄. Removal of the solvent byevaporation

gave dibenzyl 2-(3-ethoxycarbonylmethylphenyl)-2-

octanoylaminoethyl phosphite which was used for the

next reaction without further puriﬁcation. To a stirred

solution of dibenzyl 2-(3-ethoxycarbonylmethylphenyl)-

2-octanoylaminoethyl phosphite (1.58 g, 2.6 mmol) in

CH₂Cl₂(10 mL) was added m-CPBA (70%, 767 mg,

3.12 mmol) at 0 ^ꢁC and stirring was continued for 30

min at that temperature. Next saturated Na₂S₂O₃aq

was added to the mixture and the mixture was extracted

with EtOAc. The organic layer was successively washed

with saturated NaHCO₃aq, H₂O and brine before

being dried over Na₂SO₄. Removal of the solvent by

evaporation gave a residue which was puriﬁed bycol-

umn chromatographyon silica gel (Merck 7734, n-hexane/

EtOAc, 3/1–2/1–1/1) to aﬀord dibenzyl 2-(3-ethoxy-

carbonylmethylphenyl)-2-octanolyaminoethyl phosphate

as a colorless oil (80% yield in 2 steps). A mixture of

dibenzyl 2-(3-ethoxycarbonylmethylphenyl)-2-octano-

lyaminoethyl phosphate (1.3 g, 2 mmol) in EtOH (10

mL) and 10% Pd-C (130 mg) was stirred at room tem-

perature under an atmospheric pressure of hydrogen for

20 h. Removal of the catalyst by ﬁltration through a pad

of Celite followed byevaporation aﬀorded 2-(3-ethox-

ycarbonylmethylphenyl)-2-octanolyamino phosphate as

a colorless oil (89% yield). To a stirred solution of 2-(3-

ethoxycarbonylmethylphenyl)-2-octanolyamino dihydro-

gen phosphate (390 mg, 0.87 mmol) in EtOH (5 mL)

was added 1 M NaHCO₃aq. Removal of the solvent by

evaporation gave the title compound 39 as a white

powder: 94% yield; TLC R_f=0.50 (CHCl₃/MeOH/

H₂O, 65/35/8); IR (KBr) 3258, 1765, 1625, 1557, 1461,

sivelywashed with saturated NaHCO aq and brine

3

1

1025, 1098 cm^ꢀ1; H NMR (300 MHz, CD₃OD) d 7.19

before being dried over MgSO₄. Removal of the solvent

byevaporation gave an residue which was puriﬁed by

column chromatographyon silica gal (Merck 7734, n-

hexane/EtOAc, 4/1) to aﬀord 73 as a pale yellow oil:

93% yield; TLC R_f=0.35 (n-hexane/EtOAc, 3/1); ¹H

NMR (200 MHz, CDCl₃) d 8.00 (brs, 1H), 7.93 (brd,

J=7.6 Hz, 1H), 7.50 (brd, J=7.6 Hz, 1H), 7.38 (t, J=7.6

Hz, 1H), 6.30 (brd, J=7.6 Hz, 1H), 5.12–5.04 (m, 1H),

3.92 (dd, J=10.0, 4.4 Hz, 1H), 3.91 (s, 3H), 3.80 (dd,

J=10.0, 4.0 Hz, 1H), 2.25 (t, J=7.2 Hz, 2H), 2.30–2.20

(m, 2H), 1.40–1.20 (m, 8H), 0.95–0.90 (m, 12H), ꢀ0.04 (s,

3H), ꢀ0.08 (s, 3H).

(t, J=7.8 Hz, 1H), 6.98 (d, J=7.8 Hz, 1H), 6.94 (t,

J=2.1 Hz, 1H), 6.76 (m, 1H), 4.85–4.83 (m, 1H), 4.67

(s, 2H), 4.24 (q, J=7.2 Hz, 2H), 4.04–3.86 (m, 2H),

2.40–2.18 (m, 2H), 1.65–1.55 (m, 2H), 1.40–1.20 (m) and

1.28 (t, J=7.2 Hz) total 11H, 0.89 (brt, J=6.9 Hz, 3H);

MS (FAB. Pos.) m/z 512 (M+Na)⁺, 490 (M+H)⁺,

468; HRMS (MALDI-TOF, Pos.) calcd for

+

.

C₂₀H₃₁NO₈P 2Na+H : 490.1583; found: 490.1567.

1-(tert-Butyldimethylsilyl)-2-(3-methoxycarbonylphenyl)-

2-octanoylaminoethyl ether (73). To a stirred solution of

71m (10.0 g, 27.0 mmol) and TBDMSCl (4.88 g, 32.4

mmol) in DMF (30 mL) was added imidazole (4.59 g,

67.5 mmol) at 0 ^ꢁC and stirring was continued for 18 h

at room temperature. The reaction mixture was poured

N-(2-Hydroxy-1-{3-[(methoxymethoxy)methyl]phenyl}

ethyl)octanamide (74). To a stirred solution of 73 (3.00

g, 6.88 mmol) in THF (10 mL) and MeOH (20 mL) was

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3779

added LiBH₄(300 mg, 13.8 mmol) at 0 ^ꢁC and stirring

was continued at room temperature for 20 h. Next

LiBH₄(150 mg, 6.88 mmol) was added to the resulting

mixture and stirred at 45 ^ꢁC for 2 h. The reaction mix-

ture was poured into saturated NH₄Cl aq and extracted

with EtOAc. The organic layer was washed with brine

and dried over MgSO_4.Removal of the solvent byeva-

poration gave a residue which was puriﬁed bycolumn

chromatographyon silica gel (Merck 7734, n-hexane/

EtOAc, 1/1) to aﬀord 1-(tert-butyldimethylsilyl)-2-(3-

hydroxymethlyphenyl)-2-octanoylaminoethyl ether as a

colorless oil (1.80 g, 62%): TLC R_f=0.53 (n-hexane/

EtOAc, 1/1). To a stirred solution of 1-(tert-butyldi-

methylsilyl)-2-(3-hydroxymethlyphenyl)-2-octanoylamino-

that temperature. Next satd Na₂S₂O₃aq was added and

the reaction mixture was extracted with EtOAc. The

organic layer was successively washed with saturated

NaHCO₃aq, H₂O and brine before being dried over

Na₂SO₄. Removal of the solvent byevaporation gave

bis(2-cyanoethyl) 2-(3-methoxymethyloxymethylphenyl)-

2-octanoylaminoethyl phosphate as a pale yellow oil

which was used for the next reaction without further

puriﬁcation: TLC R_f=0.10 (n-hexane/EtOAc, 1/3). To a

stirred solution of bis(2-cyanoethyl) 2-(3-methoxy-

methyloxymethylphenyl)-2-octanoylaminoethyl phos-

phate in EtOH (10 mL) was added 50% Me₂NH aq (5.0

mL). Stirring was continued under reﬂux for 4 h. The

reaction mixture was poured into 1 M HCl and extrac-

ted with EtOAc. The organic layer was successively

washed with H₂O and brine before being dried over

Na₂SO₄. Removal of the solvent byevaporation gave a

residue, which was puriﬁed bycolumn chromatography

on silica gel (Merck 7734, CHCl₃/MeOH/NH₄OH, 65/

25/1–CHCl₃/MeOH/H₂O, 65/25/4) to aﬀord 2-(3-meth-

oxymethyloxymethylphenyl)-2-octanoylaminoethyl dia-

mmonium phosphate which was acidiﬁed with 1 M HCl

and then extracted with EtOAc. The organic layer was

i

ethyl ether (1.42 g, 3.38 mmol) and Pr₂NEt (3.0 mL) in

CH₂Cl₂(5.0 mL) was added MOMCl (0.385 mL, 5.07

mmol) at room temperature and stirring was continued

at that temperature for 1 h. The reaction mixture was

poured into 1 M HCl and extracted with EtOAc. The

organic layer was successively washed with saturated

NaHCO₃aq and brine before being dried over MgSO_4.

Removal of the solvent byevaporation gave 1-( tert-

butyldimethylsilyl)-2-(3-methoxymethlyoxymethylphenyl)-

2-octanoylaminoethyl ether as a yellow oil (1.39 g,

successivelywashed with H O and brine before being

2

1

89%): TLC R_f=0.31 (n-hexane/EtOAc, 3/1); H NMR

dried over Na₂SO₄. Removal of the solvent byeva-

poration gave a residue which was washed with

Et₂O/n-hexane to obtain 76 as a white powder (473 mg,

38%): TLC R_f=0.40 (CHCl₃/MeOH/H₂O, 65/35/8); ¹H

NMR (200 MHz, CD₃OD) d 7.37–7.25 (m, 4H), 5.21–

5.17 (m, 1H), 4.68 (s, 2H), 4.57 (s, 2H), 4.18–4.05 (m,

2H), 3.37 (s, 3H), 2.26 (t, J=7.5 Hz, 2H), 1.66–1.50 (m,

2H), 1.35–1.25 (m, 8H), 0.90–0.85 (m, 3H).

(200 MHz, CDCl₃) d 7.35–7.20 (m, 4H), 6.20 (brd,

J=7.6 Hz, 1H), 5.09–5.01 (m, 1H), 4.70 (s, 2H), 4.58 (s,

2H), 3.91 (dd, J=10.2, 4.2 Hz, 1H), 3.82 (dd, J=10.2,

4.2 Hz, 1H), 2.27–2.20 (m, 2H), 1.75–1.60 (m, 2H),

1.40–1.20 (m, 8H), 0.90–0.80 (m, 12H), ꢀ0.04 (s, 3H),

ꢀ0.07 (s, 3H). To a stirred solution of 1-(tert-butyldi-

methylsilyl) - 2 - (3 - methoxymethlyoxymethylphenyl) - 2-

octanoylaminoethyl ether (1.39 g, 3.00 mmol) in THF

(10 mL) was added TBAF (1.0 M THF, 3.00 mL, 3.00

mmol) at 0 ^ꢁC and stirring was continued at that tem-

perature for 30 min. The reaction mixture was poured

into saturated NH₄Cl aq and extracted with EtOAc.

The organic layer was washed with brine and dried over

MgSO₄. Removal of the solvent byevaporation gave 74 as

a y ellow oil: TLCR_f=0.34 (EtOAc). The compound was

used for the next reaction without further puriﬁcation.

2-(3-Hydroxymethylphenyl)-2-octanoylaminoethyl disodium

phosphate (41). To a stirred solution of 76 (406 mg,

0.973 mmol) in MeOH (10 mL) was added a few drops

of 1 M HCl and stirring was continued at 60 ^ꢁC for 6 h.

The reaction mixture was poured into H₂O and then

extracted with EtOAc. The organic layer was washed

with brine and dried over Na₂SO₄. Removal of the sol-

vent byevaporation gave a residue which was washed

with Et₂O/EtOAc to aﬀord 2-(3-Hydroxymethyl-

phenyl)-2-octanoylaminoethyl dihydrogen phosphate as

a white powder (279 mg, 77%): TLC R_f=0.30 (CHCl₃/

Method E

1

2-(3-Methoxymethyloxymethylphenyl)-2-octanoylamino-

ethyl phosphate (76). To a stirred solution of 74 (1.0 g,

3.0 mmol) and tetrazole (420 mg, 6.0 mmol) in CH₃CN

(20 mL) was added bis(2-cyanoethyl) diisopropylamido-

phosphite¹⁹(819 mg, 3.30 mmol) at room temperature

and stirring was continued at that temperature for 2 h.

The reaction mixture was poured into saturated

NaHCO₃aq and extracted with EtOAc. The organic

layer was successively washed with H₂O and brine

before being dried over MgSO₄. Removal of the solvent

by evaporation gave bis(2-cyanoethyl) 2-(3-methoxy-

methyloxymethylphenyl)-2-octanoylaminoethyl phos-

phite as a colorless oil which was used for the next

reaction without further puriﬁcation: TLC R_f=0.21

(n-hexane/EtOAc, 1/2). To a stirred solution of bis(2-

MeOH/H₂O, 65/35/8); HNMR (200 MHz, CD₃OD) d

7.40–7.20 (m, 4H), 5.23–5.15 (m, 1H), 4.59 (s, 2H),

4.20–4.05 (m, 2H), 2.25 (t, J=7.4 Hz, 2H), 1.65–1.50

(m, 2H), 1.40–1.20 (m, 8H), 0.91–0.85 (m, 3H). The title

compound 41 was obtained according to the same pro-

cedures as described for the preparation of 1 from 62e:

white amorphous powder; TLC R_f=0.30 (CHCl₃/

MeOH/H₂O, 65/35/8); IR (KBr) 3254, 3074, 1641, 1560,

1115, 1100, 1020, 984 cm^ꢀ^{1 1}H NMR (300 MHz,

;

CD₃OD) d 7.34 (brs, 1H), 7.25–7.18 (m, 3H), 4.90–4.87

(m, 1H), 4.56 (s, 2H), 4.03–3.88 (m, 2H), 2.37–2.19 (m,

2H), 1.65–1.53 (m, 2H), 1.34–1.25 (m, 8H), 0.91–0.86

(m, 3H); MS (FAB, Pos.) m/z 418 (M+H)⁺, 440, 396;

.

HRMS (MALDI-TOF, Pos.) calcd for C₁₇H₂₆NO₆P 2-

Na+H⁺: 418.1371; found: 418.1342.

cyanoethyl)

2-(3-methoxymethyloxymethylphenyl)-2-

octanoylaminoethyl phosphite (1.58 g, 3.0 mmol) in

CH₂Cl₂(15 mL) was added m-CPBA (57%, 908 mg, 3.0

mmol) at 0 ^ꢁC and stirring was continued for 30 min at

2-(3-Methoxymethylphenyl)-2-octanoylaminoethyl dihydro-

gen phosphate (77). To a stirred solution of 1-(tert-

butyldimethylsilyl)-2-(3-hydroxymethylyphenyl)-2-octa-

3780

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

noylaminoethyl ether (800 mg, 1.91 mmol) and MeI

(0.240 mL, 3.82 mmol) in THF (5.0 mL) was added

NaH (60% dispersion in mineral oil, 92 mg, 2.29 mmol)

at 0 ^ꢁC and stirring was continued for 2 h. The reaction

mixture was poured into saturated NH₄Cl aq and

extracted with EtOAc. The organic layer was succes-

2-(3-Methoxycarbonylphenyl)-2-octanoylaminoethyl dis-

odium phosphate (44). Bis(2-cyanoethyl) 2-(3-methoxy-

carbonylphenyl)-2-octanolyaminoethyl phosphate was

obtained from 78 according to the essentiallysame pro-

cedures as described for the preparation of 76 from 74

using DBU instead of 50% Me₂NH aq. To a stirred

sivelywashed with brine and dried over MgSO

Removal of the solvent byevaporation gave a residue

which was puriﬁed bycolumn chromatographyon silica

.

solution

of

bis(2-cyanoethyl)

2-(3-methoxy-

4

carbonylphenyl)-2-octanolyaminoethyl phosphate (2.18

mmol) in THF (10 mL) was added DBU (0.98 mL, 6.54

mmol) and stirring was continued at the reﬂux tem-

perature for 3 h. The reaction mixture was poured into

1 M HCl and extracted with EtOAc. The organic layer

was washed with brine and then dried over Na₂SO₄.

Removal of the solvent byevaporation gave a residue,

which was puriﬁed bycolumn chromatographyon silica

gel (Merck 7734, CHCl₃/MeOH/NH₄OH, 65/25/1–

CHCl₃/MeOH/H₂O, 65/25/4) to aﬀord 2-(3-methox-

ycarbonylphenyl)-2-octanoylaminoethyl diammonium

phosphate which was acidiﬁed with 1 M HCl and

extracted with EtOAc. The organic layer was washed

with brine and dried over Na₂SO₄. Removal of the sol-

vent byevaporation gave a residue which was washed

with Et₂O/n-hexane to obtain 2-(3-methoxy-

gel (Merck 7734, n-hexane/EtOAc, 4/1) to aﬀord 1-(tert-

butyldimethylsilyl)-2-(3-methoxymethylphenyl)-2-octa-

noylaminoethyl ether as a colorless oil (740 mg, 89%):

TLC R_f=0.38 (n-hexane/EtOAc, 3/1); ¹H NMR

(200 MHz, CDCl₃) d 7.35–7.20 (m, 4H), 6.20 (brd,

J=7.6 Hz, 1H), 5.09–5.01 (m, 1H), 4.44 (s, 2H), 3.90

(dd, J=10.2, 4.2 Hz, 1H), 3.80 (dd, J=10.2, 4.2 Hz,

1H), 3.41 (s, 3H), 2.27–2.10 (m, 2H), 1.75–1.60 (m, 2H),

1.40–1.20 (m, 8H), 0.90–0.80 (m, 12H), ꢀ0.04 (s, 3H),

ꢀ0.07 (s, 3H). N-{2-Hydroxy-1-[3-(methoxymethyl)

phenyl]ethyl}octanamide 75 was obtained from 1-(tert-

butyldimethylsilyl)-2-(3-methoxymethylphenyl)-2-oct-

anoylaminoethyl ether according to the same procedures

as described for the preparation of 74 from 1-(tert-butyl-

dimethylsilyl)-2-(3-methoxymethyloxymethylphenyl)-2-

octanoylaminoethyl ether, which was used for the next

reaction without further puriﬁcation: 85% yield; white

powder; TLC R_f=0.34 (EtOAc). The title compound 77

was obtained from 75 according to the same procedures

as described for the preparation of 76 from 74: 65%

yield; colorless amorphous powder; TLC R_f=0.41

(CHCl₃/MeOH/H₂O, 65/35/8); ¹H NMR (200 MHz,

CD₃OD) d 7.40–7.20 (m, 4H), 5.23–5.16 (m, 1H), 4.45

(s, 2H), 4.20–4.00 (m, 2H), 3.36 (s, 3H), 2.29–2.22 (m,

2H), 1.70–1.50 (m, 2H), 1.40–1.20 (m, 8H), 0.91–0.85

(m, 3H).

carbonylphenyl)-2-octanoylaminoethyl

dihydrogen

phosphate as a white powder (370 mg, 38%): TLC

R_f=0.41 (CHCl₃/MeOH/H₂O, 65/35/8); ¹H NMR;

(200 MHz, CD₃OD) d 8.04 (brs, 1H), 7,94 (brd, J=7.6

Hz, 1H), 7.62 (brd, J=7.6 Hz, 1H), 7.46 (t, J=7.6 Hz,

1H), 5.24 (t, J=6.2 Hz, 1H), 4.19–4.12 (m, 2H), 3.90 (s,

3H), 2.27 (t, J=5.8 Hz, 2H), 1.70–1.50 (m, 2H), 1.35–

1.20 (m, 8H), 0.90–0.84 (m, 3H). The title compound 44

was obtained from 2-(3-methoxycarbonylphenyl)-2-

octanoylaminoethyl dihydrogen phosphate according to

the same procedures as described for the preparation of

1 from 62e: white amorphous powder: TLC R_f=0.34

(CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3267, 1729,

2-(3-Methoxymethylphenyl)-2-octanoylaminoethyl diso-

dium phosphate (42). The title compound 42 was

obtained from 27 according to the same procedures as

described for the preparation of 1 from 62e: white

amorphous powder; TLC R_f=0.41 (CHCl₃/MeOH/

H₂O, 65/35/8); IR (KBr) 3260, 1626, 1555, 1102, 989

1626, 1552, 1292, 1202, 1097 cm^ꢀ^{1 1}H NMR;

;

(300 MHz, CD₃OD) d 8.01 (brs, 1H), 7.86 (brd, J=8.1

Hz, 1H), 7.61 (brd, J=8.1 Hz, 1H), 7.39 (t, J=8.1 Hz,

1H), 4.94 (dd, J=8.1, 3.9 Hz, 1H), 4.06–3.93 (m, 2H),

3.88 (s, 3H), 2.39–2.20 (m, 2H), 1.66–1.53 (m, 2H),

1.34–1.22 (m, 8H), 0.91–0.84 (m, 3H); MS (FAB, Pos.)

m/z 446 (M+H)⁺, 468, 424; HRMS (MALDI-TOF,

.

Pos.) calcd for C₁₈H₂₆NO₇P 2Na+H : 446.1321;

found: 446.1318.

1

cm^ꢀ1; H NMR (300 MHz, CD₃OD) d 7.32 (brs, 1H),

+

7.31–7.23 (m, 2H), 7.17 (brd, J=6.6 Hz, 1H), 4.92–4.87

(m, 1H), 4.42 (s, 2H), 4.02–3.88 (m, 2H), 3.33 (s, 3H),

2.38–2.19 (m, 2H), 1.66–1.53 (m, 2H), 1.35–1.24 (m,

8H), 0.92–0.85 (m, 3H); MS (FAB, Pos.) m/z 432

Trisodium 3-[1-(octanoylamino)-2-(phosphonooxy)ethyl]-

benzoate (43). To a stirred solution of 2-(3-methoxy-

carbonylphenyl)-2-octanoylaminoethyl

(M+H)⁺, 454, 410; HRMS (MALDI-TOF, Pos.)

+

.

calcd for C₁₈H₂₈NO₆P 2Na+H : 432.1479; found:

432.1528.

dihydrogen

phosphate (700 mg, 1.74 mmol) in THF (5.0 mL) and

MeOH (5.0 mL) was added 2 M NaOH (3.48 mL, 6.96

mmol) at 0 ^ꢁC and stirring was continued at room tem-

perature for 3 h. The reaction mixture was poured into

1 M HCl and extracted with EtOAc. The organic layer

was washed with brine and then dried over Na₂SO₄.

Removal of the solvent byevaporation gave a residue

which was washed with Et₂O/n-hexane to aﬀord 2-(3-

carboxylphenyl)-2-octanoylaminoethyl phosphate as a

white powder (600 mg, 89%): TLC R_f=0.20 (CHCl₃/

MeOH/H₂O, 65/35/8); ¹H NMR; (200 MHz, CD₃OD) d

8.05 (brs, 1H), 7.97–7.92 (m, 1H), 7.64–7.57 (m, 1H),

7.49–7.42 (m, 1H), 5.28–5.21 (m, 1H), 4.19–4.12 (m,

2H), 2.27 (t, J=7.6 Hz, 2H), 1.70–1.50 (m, 2H),

Methyl 3-[2-hydroxy-1-(octanoylamino)ethyl]benzoate (78).

The title compound 78 was obtained from 73 according

to the same procedures as described for the preparation

of 74 from 1-(tert-butyldimethylsilyl)-2-(3-methoxy-

methlyoxy methylphenyl)-2-octanoylaminoethyl ether,

which was washed with n-hexane to give a white powder:

93% yield; TLC R_f=0.25 (n-hexane/EtOAc, 1/3); ¹H

NMR (300 MHz, CDCl₃) d 8.00–7.94 (m, 2H), 7.51 (dt,

J=6.0 Hz, 1.5 Hz, 1H), 7.43 (t, J=8.1 Hz, 1H), 6.26 (d,

J=6.9 Hz, 1H), 5.15–5.09 (m, 1H), 3.91 (s, 3H), 3.92–

3.87 (m, 2H), 2.26 (t, J=7.5 Hz, 2H), 1.67–1.62 (m, 2H),

1.30–1.26 (m, 8H), 0.86(t, J=6.6 Hz, 3H).

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3781

1.40–1.20 (m, 8H), 0.90–0.84 (m, 3H). The title com-

pound 43 was prepared from 2-(3-carboxylphenyl)-2-

octanoylaminoethyl dihydrogen phosphate according to

the same procedures as described for the preparation of

1 from 62e: white amorphous powder: TLC R_f=0.20

(CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3422, 1637,

was washed with brine, dried over Na₂SO₄and con-

centrated to aﬀord a residue, which was puriﬁed by

column chromatographyon silica gel (Merck 7734,

n-hexane/EtOAc, 8/2–6/4) to aﬀord a diazoketone deri-

1

vative: TLC R_f=0.48 (n-hexane/EtOAc, 1/1); H NMR

(300 MHz, CDCl₃) d 7.74 (t, J=1.5 Hz, 1H), 7.62 (dt,

J=7.8, 1.5 Hz, 1H), 7.51–7.48 (m, 1H), 7.39 (t, J=7.8

Hz, 1H), 7.33–7.22 (m, 5H), 6.26 (d, J=7.8 Hz, 1H),

5.83 (s, 1H), 4.53 (d, J=11.7 Hz, 1H), 4.48 (d, J=11.7

Hz, 1H), 3.78–3.69 (m, 2H), 2.2 1(t, J=7.2 Hz, 1H),

1.65–1.56 (m, 2H), 1.28–1.23 (m, 8H), 0.86 (t, J=6.9

Hz, 3H). To a stirred solution of the above-described

diazoketone (1.53 g, 3.63 mmol) and Et₃N (0.52 mL) in

MeOH (36 mL) was added silver acetate (121 mg, 7.27

mmol) and stirring was continued at 40 ^ꢁC for 1 h. The

reaction mixture was diluted with EtOAc. After the

organic layer was successively washed with 1 M HCl

and brine, it was dried over Na₂SO₄. Removal of the

solvent byevaporation gave a residue which was pur-

iﬁed bycolumn chromatographyon silica gel (Merck

7734, n-hexane/EtOAc, 7/3–6/4) to aﬀord 79 (1.25 g,

49% in 3 steps): TLC R_f=0.53 (n-hexane/EtOAc, 1/1);

¹H NMR (300 MHz, CDCl₃) d 7.35–7.17 (m, 9H), 6.15

(d, J=7.8 Hz, 1H), 5.23–5.18 (m, 1H), 4.53 (d, J=12.0

Hz, 1H), 4.48 (d, J=12.0 Hz, 1H), 3.75 (dd, J=9.9, 4.8

Hz, 1H), 3.71 (d, J=9.9, 4.8 Hz, 1H), 3.67 (s, 3H), 3.60

(s, 2H), 2.23–2.18 (m, 2H), 1.68–1.57 (m, 2H), 1.29–1.24

(m, 8H), 0.87 (t, J=6.6 Hz, 3H).

1560, 1390, 1096, 984 cm^ꢀ^{1 1}H NMR; (300 MHz,

;

CD₃OD) d 7.96 (brs, 1H), 7.80 (dt, J=7.5, 1.2 Hz, 1H),

7.42 (brd, J=7.5 Hz, 1H), 7.26 (t, J=7.5 Hz, 1H), 4.98–

4.94 (m, 1H), 4.04–3.90 (m, 2H), 2.38–2.20 (m, 2H),

1.65–1.54 (m, 2H), 1.34–1.22 (m, 8H), 0.90–0.86 (m,

3H); MS (FAB, Pos.) m/z 454 (M+H)⁺, 432; HRMS

(MALDI-TOF, Pos.) calcd for disodium salt

+

.

C₁₇H₂₄NO₇P 2Na+H : 432.1164; found: 432.1134.

3-[2-(Benzyloxy)-1-(octanoylamino)ethyl]benzoic acid (79).

To a stirred mixture of 78 (7.41 g, 23.08 mmol) and

Ag₂O (6.42 g, 27.7 mmol) in DMF (23 mL) was added

benzyl bromide (3.29 mL, 27.7 mmol) at room tem-

perature and stirring was continued for 20 h at that

temperature. The reaction mixture was poured into H₂O

and extracted with EtOAc. The organic layer was

washed with brine and dried over Na₂SO₄. Removal of

the solvent byevaporation gave a residue which was

puriﬁed bycolumn chromatographyon silica gel

(Merck 7734, n-hexane/EtOAc, 85/15) to aﬀord methyl

3-[2-(benzyloxy)-1-(octanoylamino)ethyl]benzoate, which

was used for the next reaction without further puriﬁ-

cation: TLC R_f=0.39 (n-hexane/EtOAc, 4/1). To a stirred

solution

of

methyl

3-[2-(benzyloxy)-1-(octanoyl-

2-(3-Methoxyarboxylmethylphenyl)-2-octanoylaminoethyl

disodium phosphate (46). A mixture of 80 (1.21 g, 2.85

mmol) and 10% Pd–C (1 g) was vigorouslystirred at

room temperature under an atmospheric pressure of

hydrogen for 2 h. Removal of the catalyst by ﬁltration

through a pad of Celite followed byevaporation aﬀor-

ded 2-(3-methoxyarboxylmethylphenyl)-2-octanoylamino-

ethanol as an oilyresidue which was used for the next

reaction without further puriﬁcation: TLC R_f=0.21

amino)ethyl]benzoate (23.0 mmol) in THF (2 mL) and

MeOH (14 mL) was added 2 M NaOH (14.8 mL, 29.6

mmol) at 0 ^ꢁC and stirring was continued for 2 h at

room temperature. The reaction mixture was poured

into H₂O and extracted with Et₂O. The aqueous layer

was acidiﬁed with 2 M HCl and extracted with EtOAc.

The organic layer was washed with brine and dried over

Na₂SO₄. Removal of the solvent byevaporation gave a

residue which was washed with Et₂O/n-hexane to aﬀord

79 (89 g, 53% yield in 2 steps): TLC R_f=0.50 (CHCl₃/

1

(n-hexane/EtOAc, 1/3); H NMR (300 MHz, CDCl₃) d

7.32 (t, J=7.5 Hz, 1H), 7.23–7.18 (m, 3H), 6.20 (d,

J=5.7 Hz, 1H), 5.08–5.00 (m, 1H), 3.87–3.85 (m, 2H),

3.69 (s, 3H), 3.62 (s, 2H), 2.28–2.19 (m, 2H), 1.71–1.60

(m, 2H), 1.29–1.23 (m, 8H), 0.87 (t, J=6.3 Hz, 3H). The

title compound 46 was prepared from 2-(3-methoxy-

carboxylmethylphenyl)-2-octanoylaminoethanol accord-

ing to the same procedure as described in the

preparation of 39 from 72: ivorypowder; TLC R_f=0.42

(CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3862, 3423,

2928, 2857, 1739, 1637, 1550, 1491, 1438, 1345, 1258,

1

MeOH, 9/1); H NMR (300 MHz, CDCl₃) d 8.08–8.06

(m, 1H), 7.99 (dt, J=7.8, 1.5 Hz, 1H), 7.58–7.54 (m,

1H), 7.42 (t, J=7.2 Hz, 1H), 7.36–7.23 (m, 5H), 6.31 (d,

J=7.2 Hz, 1H), 5.27–5.22 (m, 1H), 4.54 (d, J=12.0 Hz,

1H), 4.49 (d, J=12.0 Hz, 1H), 3.78 (dd, J=9.9, 4.5 Hz,

1H), 3.73 (dd, J=9.9, 4.5 Hz, 1H), 2.24 (t, J=7.5 Hz,

2H), 1.69–1.58 (m, 2H), 1.36–1.23 (m, 8H), 0.86 (t,

J=7.3 Hz, 3H).

1

Methyl {3-[2-(benzyloxy)-1-(octanoylamino)ethyl]phenyl}

acetate (80). To a stirred solution of 79 (2.38 g, 6.0

mmol) in toluene (60 mL) and DMF (0.1 mL) was

added dropwise (COCl)₂(0.63 mL, 7.2 mmol) at 0 ^ꢁC

and stirring was continued at room temperature for 2 h.

Removal of the solvent byevaporation gave an acid

chloride which was used for the next reaction without

further puriﬁcation: TLC R_f=0.33 (n-hexane/EtOAc, 2/

1). To a stirred mixture of the acid chloride and

NaHCO₃(504 mg, 6.0 mmol) in Et₂O (60 mL) was

added CH₂N₂/Et₂O(100 mL) at 0 ^ꢁC and stirring was

continued at that temperature for 2 h. Removal of the

solvent byevaporation gave a residue which was poured

into H₂O and extracted with Et₂O. The organic layer

1091, cm^ꢀ1; H NMR (300 MHz, CD₃OD) d 7.25–7.19

(m, 3H), 7.13–7.10 (m, 1H), 4.90–4.85 (m, 1H), 4.02–

3.87 (m, 2H), 3.65 (s, 3H), 3.60 (s, 2H), 2.37–2.18 (m,

2H), 1.63–1.55 (m, 2H), 1.37–1.25 (m, 8H), 0.88 (t,

J=6.9 Hz, 3H); MS (FAB. Pos.) m/z 460 (M+H)⁺;

.

HRMS (MALDI-TOF, Pos.) calcd for C₁₉H₂₈NO₇P 2-

Na+H⁺: 460.1477; found: 460.1432.

Trisodium 2-(3-carboxymethlyphenyl)-2-octanoylamino-

ethyl phosphate (45). To a stirred solution of 2-(3-meth-

oxycarboxylmethylphenyl)-2-octanoylaminoethyl dihy-

drogen phosphate (490 mg, 1.07 mmol) in THF (3.0

mL) and MeOH (3.0 mL) was added 1 M NaOH (1.61

mL, 1.61 mmol) and stirring was continued at that

3782

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

temperature for 20 h. The reaction mixture was poured

into 1 M HCl and extracted with EtOAc–MeOH. The

organic layer was washed with brine and dried over

Na₂SO_4.Removal of the solvent byevaporation gave a

residue which was puriﬁed bycolumn chromatography

on silica gel (Merck 7734, CHCl₃/MeOH/NH₄OH, 65/

25/1–CHCl₃/MeOH/H₂O, 65/35/8) to aﬀord 2-(3-car-

boxyphenyl)-2-octanoylaminoethyl dihydrogen phos-

phate as a white amorphous powder (208 mg, 48%):

carbonyl]thio}phenyl) (octanoylamino)acetate (1.62 g,

4.1 mmol) in THF (15 mL) was added LiBH₄(179 mg,

8.2 mmol) and stirring was continued at room tempera-

ture for 2 h. The reaction was quenched with saturated

NH₄Cl aq and extracted with EtOAc. The organic layer

was washed with brine and dried over Na₂SO₄.

Removal of the solvent byevaporation gave 2-(3-dime-

thycarbonylthiophenyl)-2-octanoylaminoethanol which

was used for the next reaction without further puriﬁ-

cation: TLC R_f=0.16 (toluene/EtOAc, 3/1); MS

(MALDI-TOF, Pos.) m/z 389 (M+Na)⁺, 367

(M+H)⁺; ¹H NMR (200 MHz, CDCl₃) d 7.40–7.26 (m,

4H), 6.37 (brd, J=7.2 Hz, 1H), 5.04 (m, 1H), 3.86 (dd,

J=11.6, 4.6 Hz, 1H), 3.79 (dd, J=11.6, 4.0 Hz, 1H),

3.20–2.95 (brs, 6H), 2.24 (brt, J=7.6 Hz, 2H), 1.70–1.55

(m, 2H), 1.40–1.20 (m, 8H), 0.87 (brt, J=6.8 Hz, 3H).

To a stirred solution of 2-(3-dimethycarbonylthiophe-

nyl)-2-octanoylaminoethanol (4.1 mmol) in MeOH (20

mL) was added KOH (2.0 g, 35.6 mmol) and stirring

was continued under reﬂux for 40 min. The reaction

mixture was poured into 1 M HCl (50 mL) and extrac-

ted with EtOAc. The organic layer was washed with

brine and then dried over Na₂SO₄. Removal of the sol-

vent byevaporation gave a solid which was washed with

Et₂O/n-hexane to aﬀord 2-(3-mercaptophenyl)-2-octa-

noylaminoethanol (820 mg, 67%): TLC R_f=0.19 (n-

hexane/EtOAc, 2/3); MS (MALDI-TOF, Pos.) m/z 318

(M+Na)⁺, 296 (M+H)⁺; ¹H NMR (300 MHz,

CDCl₃) d 7.20–7.15 (m, 4H), 6.25 (brd, J=7.2 Hz, 1H),

5.00 (m, 1H), 3.84 (m, 2H), 2.22 (brt, J=7.6 Hz, 2H),

1.70–1.55 (m, 2H), 1.40–1.20 (m, 8H), 0.86 (brt, J=6.8

Hz, 3H). To a stirred mixture of 2-(3-mercaptophenyl)-

2-octanoylaminoethanol (590 mg, 2 mmol) and K₂CO₃

(691 mg, 5 mmol) was added 2-idopropane (510 mg, 3

mmol) and stirring was continued at 70 ^ꢁC for 3 h. After

cooling, the precipitates were removed byﬁltration

through a pad of Celite. Evaporation of the ﬁltrate gave

an oilyresidue, which was dissolved in EtOAc. The

organic layer was successively washed with H₂O and brine

before being dried over MgSO₄and concentrated to give

an oilyresidue, which was puriﬁed bycolumn chromato-

graphyon silica gel (Merck 7734, n-hexane/EtOAc, 1/1–2/

3) to aﬀord 81 (654 mg, 97%): TLC R_f=0.38 (n-hexane/

EtOAc, 2/3); MS (MALDI-TOF, Pos.) m/z 376

(M+K)⁺, 360 (M+Na)⁺, 338 (M+H)⁺; ¹H NMR

(300MHz, CDCl₃) d 7.33–7.26 (m, 3H), 7.12 (m, 1H), 6.16

(brd, J=6.6 Hz, 1H), 5.03 (m, 1H), 3.86 (m, 2H), 3.38 (m,

1H), 2.69 (brs, 1H), 2.24 (brt, J=7.5 Hz, 2H), 1.70–1.55

(m, 2H), 1.40–1.20 (m, 8H), 0.87 (brt, J=6.6 Hz, 3H).

1

TLC R_f=0.28 (CHCl₃/MeOH/H₂O, 65/35/8); H NMR

(300 MHz, CD₃OD) d 7.33–7.15 (m, 4H), 5.21–5.16 (m,

1H), 4.19–4.06 (m, 2H), 3.60 (s, 2H), 2.28–2.23 (m, 2H),

1.66–1.54 (m, 2H), 1.35–1.24 (m, 8H), 0.91–0.86 (m, 3H).

The title compound 45 was obtained from 2-(3-carboxy-

phenyl)-2-octanoylaminoethyl phosphate according to

essentiallythe same procedure as described for the pre-

paration of 1 from 62e: white powder: TLC R_f=0.28

(CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3423, 1637,

1577, 1388, 1093, 984 cm^ꢀ^{1 1}H NMR; (300 MHz,

;

CD₃OD) d 7.26 (brs, 1H), 7.18 (brs, 3H), 4.92–4.88 (m,

1H), 4.02–3.88 (m, 2H), 3.45 (s, 2H), 2.34–2.18 (m, 2H),

1.66–1.54 (m, 2H), 1.36–1.24 (m, 8H), 0.90–0.86 (m,

3H); MS (FAB, Pos.) m/z 468 (M+H)⁺, 446, 424;

.

HRMS (MALDI-TOF, Pos.) calcd for C₁₈H₂₅NO₇P 3-

Na+H⁺: 468.1140; found: 468.1102.

N-{2-Hydroxy-1-[3-(isopropylthio)phenyl]ethyl}octanamide

(81). To a stirred solution of 70 (3.07 g, 10 mmol) in

DMF (15 mL) was added NaH (60% dispersion in

mineral oil, 420 mg, 10 mmol) at 0 ^ꢁC. Dimethylthio-

carbamoyl chloride (1.61 g, 13 mmol) was then added to

the resulting mixture and stirring was continued at

80 ^ꢁC for 1.5 h. The reaction mixture was poured into

H₂O and extracted with EtOAc. The organic layer was

successivelywashed with 1 M HCl, saturated NaHCO

3

aq and brine before being dried over Na₂SO₄. Removal

of the solvent byevaporation gave a residue which was

puriﬁed bycolumn chromatographyon silica gel

(Merck 7734, toluene/EtOAc, 7/1–5/1) and (FL60D

toluene/EtOAc, 10/1) to aﬀord methyl (3-{[(dimethyl-

amino)carbonothioyl]oxy}phenyl) (octanoylamino)acetate

(2.55 g, 64%): TLC R_f=0.37 (toluene/EtOAc, 3/1); MS

(MALDI-TOF, Pos.) m/z 417 (M+Na)⁺, 395

(M+H)⁺; ¹H NMR (300 MHz, CDCl₃) d 7.37 (dd,

J=7.8, 7.8 Hz, 1H), 7.28–7.04 (m, 3H), 6.40 (brd,

J=6.9 Hz, 1H), 5.59 (d, J=6.9 Hz, 1H), 3.74 (s, 3H),

3.45 (s, 3H), 3.33 (s, 3H), 2.24 (brt, J=7.5 Hz, 2H),

1.70–1.55 (m, 2H), 1.40–1.20 (m, 8H), 0.87 (brt, J=6.9

Hz, 3H). A solution of methyl (3-{[(dimethylamino)-

carbonothioyl]oxy}phenyl) (octanoylamino)acetate (2.07

g, 5.24 mmol) in diphenyl ether (35 mL) was heated with

stirring to 235 ^ꢁC and the stirring was continued for 10

h. After cooling, the reaction mixture was puriﬁed by

column chromatographyon silica gel (Merck7734, tolu-

ene/EtOAc, 5/1–3/1) to give methyl (3-{[(dimethylamino)

carbonyl]thio}phenyl) (octanoylamino)acetate (1.62 g,

66%): TLC R_f=0.18 (toluene/EtOAc, 3/1); MS

(MALDI-TOF, Pos.) m/z 417 (M+Na)⁺, 395

(M+H)⁺; ¹H NMR (300 MHz, CDCl₃) d 7.50–7.35 (m,

4H), 6.42 (brd, J=6.9 Hz, 1H), 5.59 (d, J=6.9 Hz, 1H),

3.15–2.95 (brs, 6H), 2.24 (brt, J=7.5 Hz, 2H), 1.70–1.55

(m, 2H), 1.40–1.20 (m, 8H), 0.87 (brt, J=6.9 Hz, 3H).

To a stirred solution of methyl (3-{[(dimethylamino)-

2-(3-Isopropylthiophenyl)-2-octanoylaminoethyl disodium

phosphate (48). The title compound 48 was prepared

from 81 according to the same procedure as described

for the preparation of 22 from 60t (general method C):

oﬀ-white powder; TLC R_f=0.27 (CHCl₃/MeOH/H₂O,

65/25/4); IR (KBr) 3264, 2926, 1659, 1625, 1555, 1464,

1

1097 cm^ꢀ1; H NMR (200 MHz, CD₃OD) d 7.36 (brs,

1H), 7.22 (m, 3H), 4.84 (m, 1H), 4.00–3.90 (m, 2H), 3.37

(m, 1H), 2.41–2.17 (m, 2H), 1.70–1.50 (m, 2H), 1.40–

1.20 (m, 8H), 1.24 (d, J=6.6 Hz, 6H), 0.88 (brt, J=6.6

Hz, 3H); MS (FAB, Pos.) m/z 484 (M+Na)⁺, 462

(M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd for

C₁₉H₃₀NO₅PS^ꢃ2Na+H⁺: 462.1456; found: 462.1435.

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3783

2-(3-Methylsulfonylphenyl)-2-octanoylaminoethyl disodium

phosphate (49). 2-(3-Methylthiophenyl)-2-octanoyl-

aminoethyl dihydrogen phosphate was prepared

according to the same procedure as described for the

preparation of 47 from 64k. To a stirred solution of

2-(3-methylthiophenyl)-2-octanoylaminoethyl dihydro-

gen phosphate (320 mg, 0.823 mmol) in MeOH was

organic layer was successively washed with 1 M NaOH

and brine before being dried over Na₂SO₄. Removal of

the solvent byevaporation gave a residue which was

puriﬁed bycolumn chromatographyon silica gel (Merck

7734, n-hexane/EtOAc, 2/1) to aﬀord 2-(3-aminophenyl)-

1-(tert-butyldimethylsilyl)-2-octanonylaminoethyl ether

as a yellow oil (961 mg, 36% yield in 2 steps): TLC

added OXONE¹(2KHSO₅KHSO₄K₂SO₄, 556 mg,

0.905 mmol) at 0 ^ꢁC and stirring was continued at room

temperature for 6 h. The reaction mixture was poured

into 0.5 M HCl and extracted with EtOAc. The organic

layer was successively washed with H₂O and brine

before being dried over MgSO₄. Removal of the solvent

by evaporation gave 2-(3-methylsulfonylphenyl)-2-octa-

noylaminoethyl dihydrogen phosphate as a beige amor-

phous powder (246 mg, 71%): TLC R_f=0.31 (CHCl₃/

R_f=0.29 (n-hexane/EtOAc, 1/2); H NMR (300 MHz,

1

.

CDCl₃) d 7.10 (t, J=7.8 Hz, 1H), 6.71 (d, J=7.8 Hz,

1H), 6.66 (brs, 1H), 6.61–6.58 (m, 1H), 6.14 (d, J=7.8

Hz, 1H), 4.97–4.91 (m, 1H), 3.87 (dd, J=10.2, 4.5 Hz,

1H), 3.78 (dd, J=10.2, 4.5 Hz, 1H), 2.25–2.20 (m, 2H),

1.70–1.60 (m, 2H), 1.40–1.20 (m, 8H), 0.88–0.86 (m,

12H), ꢀ0.03 (s, 3H), ꢀ0.06 (s, 3H). To a stirred solution

of 2-(3-aminophenyl)-1-(tert-butyldimethylsilyl)-2-octa-

nonylaminoethyl ether (426 mg, 1.08 mmol) in pyridine

(5.0 mL) was added dropwise MeSO₂Cl (0.125 mL, 1.62

mmol) at 0 ^ꢁC and stirring was continued at that tem-

perature for 30 min. The reaction mixture was poured

into 1 M HCl and extracted with EtOAc. The organic

layer was washed with brine and dried over Na₂SO₄.

Removal of the solvent byevaporation gave a residue,

which was puriﬁed bycolumn chromatographyon silica

gel (Merck 7734, n-hexane/EtOAc, 1/1) to aﬀord methyl

{3 - [(methylsulfonyl)amino]phenyl}(octanoylamino)ace-

tate as a yellow oil (508 mg, quant.): TLC R_f=0.52

(n-hexane/EtOAc, 1/1); MS (APCI, Pos., 20eV) m/z 471

(M+H)⁺; ¹H NMR (200 MHz, CDCl₃) d 7.32–7.24 (m,

1H), 7.15–7.06 (m, 3H), 6.96 (brs, 1H), 6.34 (d, J=7.2

Hz, 1H), 5.03–4.95 (m, 1H), 3.90 (dd, J=10.2, 4.0 Hz,

1H), 3.78 (dd, J=10.2, 4.4 Hz, 1H), 2.94 (s, 3H), 1.80–

0.85 (m, 3H), 0.85 (s, 9H), ꢀ0.04 (s, 3H), ꢀ0.07 (s, 3H).

1

MeOH/H₂O, 65/35/8); H NMR (300 MHz, CD₃OD) d

7.97 (brs, 1H), 7.88 (brd, J=7.8 Hz, 1H), 7.72 (brd,

J=7.8 Hz, 1H), 7.62 (t, J=7.8 Hz, 1H), 5.30–5.20 (m,

1H), 4.20–4.16 (m, 2H), 3.11 (s, 3H), 2.28 (t, J=7.5 Hz,

2H), 1.65–1.55 (m, 2H), 1.35–1.20 (m, 8H), 0.90–0.85

(m, 3H). The title compound 49 was prepared from

2-(3-methylsulfonylphenyl)-2-octanoylaminoethyl phos-

phate according to the same procedure for as described

the preparation of 1 from 62e: beige amorphous pow-

der; TLC R_f=0.31(CHCl₃/MeOH/H₂O, 65/35/8); IR

(KBr) 3375, 1644, 1546, 1299, 1147, 1094, 983 cm^ꢀ1; ¹H

NMR (200 MHz, CD₃OD) d 7.95 (brs, 1H), 7.80 (brd,

J=7.8 Hz, 1H), 7.74 (brd, J=7.8 Hz, 1H), 7.55 (t,

J=7.8 Hz, 1H), 5.00–4.95 (m, 1H), 4.14–3.89 (m, 2H),

3.10 (s, 3H), 2.43–2.18 (m, 2H), 1.68–1.52 (m, 2H),

1.36–1.22 (m, 8H), 0.93–0.84 (m, 3H); MS (FAB, Pos.)

m/z 466 (M+Na)⁺, 444 (M+H)⁺, 422; HRMS

.

(MALDI-TOF, Pos.) calcd for C₁₇H₂₆NO₇PS 2

Na+H⁺: 466.1041; found: 466.1026.

Trisodium 2-{3-[(methylsulfonyl)amino]phenyl}-2-(octanoyl-

amino)ethyl phosphate (50). 3-[(Methylsulfonylamino)-

phenyl]-2-octanonylaminoethanol was prepared from

1-(tert-Butyldimethylsilyl)-2-{3-[(diphenylmethylene)amino]-

phenyl}-2-octanoylaminoethyl ether (83). To a stirred

solution of 82 (3.57 g, 6.79 mmol) and 1,1-diphe-

nylmethanimine (1.85 g, 10.2 mmol) in toluene (15 mL)

were added Cs₂CO₃(3.10 g, 9.51 mmol), Pd(OAc)₂(15

mg, 0.0679 mmol) and (ꢂ)-BINAP (64 mg, 0.102

mmol) and stirring was continued at 80 ^ꢁC for 20 h. The

reaction mixture was diluted with Et₂O. Removal of the

catalyst by ﬁltration through a pad of Celite followed by

evaporation of the ﬁltrate aﬀorded 83 as a brown oil

(6.10 g) which was used for the next reaction without

further puriﬁcation: TLC R_f=0.70 (n-hexane/EtOAc, 3/

methyl

{3-[(methylsulfonyl)amino]phenyl}(octanoyl-

amino)acetate according to the essentiallysame proce-

dure as described for the preparation of 74 from 1-(tert-

butyldimethylsilyl) - 2 - [(3 - methoxymethlyoxymethyl)-

phenyl]-2-octanoylaminoethyl ether: yellow viscous oil;

75% yield; TLC R_f=0.42 (EtOAc). The title compound

50 was prepared from 84 according to the same proce-

dure as described for the preparation of 39 from 72

(General Method D): 84% yield; white powder; TLC

R_f=0.29 (CHCl₃/MeOH/H₂O, 65/35/8); IR (KBr) 3299,

1636, 1550, 1484, 1283, 1216, 1110, 982 cm^ꢀ1; HNMR

1

(300 MHz, CD₃OD) d 7.08 (brs, 1H), 7.05 (t, J=7.5 Hz,

1H), 6.93 (brd, J=7.5 Hz, 1H), 6.79 (brd, J=7.5 Hz,

1H), 4.84–4.81 (m, 1H), 4.00–3.87 (m, 2H), 2.79 (s, 3H),

2.36–2.18 (m, 2H), 1.66–1.54 (m, 2H), 1.35–1.25 (m,

8H), 0.91–0.86 (m, 3H); MS (FAB, Pos.) m/z 503

(M+H)⁺, 481, 525; HRMS (MALDI-TOF, Pos.) calcd

.

for free acid form C₁₇H₂₉N₂O₇PS +Na : 459.1331;

found: 459.1344.

1

1); H NMR (300 MHz, CDCl₃) ꢀ 7.74–7.71 (m, 2H),

7.50–7.37 (m, 4H), 7.28–7.20 (m, 3H), 7.12–7.05 (m,

3H), 6.86 (d, J=7.8 Hz, 1H), 6.67–6.60 (m, 2H), 5.91 (d,

J=7.8 Hz, 1H), 4.91–4.86 (m, 1H), 3.74 (dd, J=10.2,

4.5 Hz, 1H), 3.60 (dd, J=10.2, 4.8 Hz, 1H), 2.25–2.20

(m, 2H), 1.70–1.60 (m, 2H), 1.40–1.20 (m, 8H), 0.88–

0.86 (m, 12H), ꢀ0.04 (s, 3H), ꢀ0.06 (s, 3H).

+

1-(tert-Butyldimethylsilyl)-2-[(3-methylsulfonylamino)-

phenyl]-2-octanonylamino ether (84). To a stirred solu-

tion of 83 (6.10 g, 6.79 mmol) in MeOH (100 mL) were

added CH₃CO₂Na (3.94 g, 48.0 mmol) and

NH₂OH HCl (2.50 g, 36.0 mmol). Stirring was con-

tinued at room temperature for 30 min. The reaction

mixture was evaporated and extracted with EtOAc. The

N-[(1R)-2-Hydroxy-1-(3-methoxyphenyl)ethyl]octanamide

(86). The compound 85 was prepared from 1-methoxy-

3-vinylbenzene according to the known procedure:²³

(>99% ee); white powder; TLC R_f=0.53 (EtOAc/

AcOH/H₂O 3/1/1); ¹H NMR (300 MHz, CDCl₃) d 8.65–

8.50 (br, 3H), 7.34–7.26 (m, 1H), 7.15 (brs, 1H), 7.04 (d,

J=7.8 Hz, 1H), 6.92 (dd, J=8.1, 2.7 Hz, 1H), 5.25 (t,

.

3784

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

J=4.8 Hz, 1H), 4.21 (t, J=6.3 Hz, 1H), 3.76 (s, 3H),

3.74–3.68 (m, 2H); ee determination: the corresponding

4-nitro benzoate prepared from 85 was analyzed by

HPLC: DAICEL CHIRALCEL OD (0.46ꢄ25 cm);

column temperature, 25 ^ꢁC; eluent, EtOH/n-hex-

ane=30/70; ﬂow rate, 0.5 mL /min; UV 260 nm; the

retention times of 4-nitro benzoate and its enantiomer

were 19.7 and 11.7 min, respectively. The title com-

pound 86 was prepared quantitativelyfrom 85 accord-

ing to essentiallythe same procedure as described for

the preparation of 60e from (2R)-2-amino-2-phenyl-

ethanol. Compound 86 was then used for the next reac-

tion without further puriﬁcation: quant.; white solid;

TLC R_f=0.40 (n-hexane/EtOAc 1/2); ¹H NMR

(300 MHz, CDCl₃) d 7.33–7.26 (m, 1H), 6.90–6.80 (m,

3H), 6.09 (d, J=6.3 Hz, 1H), 5.05–5.00 (m, 1H), 3.95–

3.82 (m, 2H), 3.81 (s, 3H), 2.75–2.60 (br, 1H), 2.28–2.21

(m, 2H), 1.70–1.50 (m, 2H), 1.40–1.20 (m, 8H), 0.88 (t,

J=6.8 Hz, 3H).

CD₃OD) d 7.17 (t, J=8.0 Hz, 1H), 6.92–6.89 (m, 2H),

6.76–6.71 (m, 1H), 4.89–4.83 (m, 1H), 3.98–3.90 (m,

2H), 3.76 (s, 3H), 2.42–2.17 (m, 2H), 1.67–1.55 (m, 2H),

1.29–1.25 (m, 8H), 0.88 (t, J=7.0 Hz, 3H); MS (FAB,

pos.) m/z 418 (M+H)⁺; HRMS (MALDI-TOF, Pos.)

calcd for C₁₇H₂₆NO₆P^ꢃ2Na+H⁺: 418.1371; found;

418.1347.

Method F

(2S)-2-Octanoylamino-2-phenylethyl disodium phosphate

(52). To a stirred solution of 89 (3.0 g, 12.7 mmol) in

pyridine (30 mL) was added bis(2,2,2-trichloroethyl)-

chloridophosphate (5.76 g, 15.2 mmol) at 0 ^ꢁC and stir-

ring was continued at that temperature for 1.5 h. The

reaction mixture was poured into ice-cold 1 M HCl and

extracted with EtOAc. The organic layer was succes-

sivelywashed with 1 M HCl, H ₂O and brine before

being dried over Na₂SO₄. Removal of the solvent by

evaporation gave a residue, which was washed with

Et₂O to aﬀord 90 as a white solid (1.14 g). The ﬁltrate

was concentrated to give a reside, which was puriﬁed by

column chromatographyon silica gel (Merck 7734, n-

hexane/EtOAc, 4/1) to aﬀord additional 90 (5.36 g). In

(2R)-2-(3-Methoxyphenyl)-2-octanoylaminoethyl disodium

phosphate (2). The title compound 2 was prepared from

86 according to the same procedure as described for the

preparation of 1 from 60e (general method A): light

brown powder, TLC R_f=0.51 (CHCl₃/MeOH/H₂O 65/

35/8); IR (KBr) 3399, 2956, 2929, 2858, 1638, 1548,

1492, 1466, 1437, 1262, 1092 cm^ꢀ1; ¹H NMR (300 MHz,

CD₃OD) d 7.18 (dd, J=8.1, 8.1 Hz, 1H), 6.95–6.88 (m,

2H), 6.78–6.72 (m, 1H), 4.95–4.80 (m, 1H), 4.02–3.85

(m, 2H), 3.77 (s, 3H), 2.40–2.20 (m, 2H), 1.70–1.50 (m,

2H), 1.40–1.20 (m, 8H), 0.89 (t, J=6.8 Hz, 3H); optical

rotation [a]²_D⁵-43.82 (c 1.01, H₂O); MS (FAB, pos.) m/z

1

total 6.5 g (89%) of compound 90 was obtained: H

NMR (200 MHz, CDCl₃) d 7.42–7.20 (m, 5H), 5.28 (m,

1H), 5.03 (m, 1H), 4.60–4.30 (m, 6H), 1.43 (s, 9H). To a

solution of 90 (4.23 g, 7.33 mmol) in CH₂Cl₂(20 mL)

was added triﬂuoroacetic acid (10 mL) at 0 ^ꢁC and stir-

ring was continued for 1 h at room temperature.

Removal of the solvent byevaporation gave (2 S)-2-

amino-2-phenylethyl bis(2,2,2-trichloroethyl) phosphate

hydrochloride which was used for the next reaction

without further puriﬁcation. To a stirred solution of

(2S)-2-amino-2-phenylethyl bis(2,2,2-trichloroethyl) phos-

phate hydrochloride (7.33 mmol) in CH₂Cl₂(10 mL)

were added octanoyl chloride (1.43 g, 8.8 mmol) and

pyridine (1.78 mL, 22 mmol) at 0 ^ꢁC and stirring was

continued at room temperature for 15 min. Removal of

the solvent byevaporation gave (2 S)-2-phenyl-2-(octa-

noylamino)ethyl bis(2,2,2-trichloroethyl) phosphate

which was used for the next reaction without further

puriﬁcation. To a stirred solution of (2S)-2-phenyl-2-

(octanoylamino)ethyl bis(2,2,2-trichloroethyl) phos-

phate (7.33 mmol) in pyridine (25 mL)/AcOH (5 mL)

was added Zn powder (4.2 g, 64.5 mmol) at 0 ^ꢁC and

stirring was continued at room temperature for 2.5 h.

Removal of the Zn powder byﬁltration through a pad

of Celite followed byevaporation aﬀorded an oilyresi-

due, which was dissolved in 5 M NaOH and extracted

with EtOAc. The aqueous layer was acidiﬁed by 2 M

HCl and extracted with EtOAc. The organic layer was

440 (M+Na)⁺, 418 (M+H)⁺, 396; HRMS (MALDI-

+

.

TOF, Pos.) calcd for C₁₇H₂₆NO₆P 2Na+H : 418.1371;

found; 418.1413.

N-[(1S)-2-Hydroxy-1-(3-methoxyphenyl)ethyl]octanamide

(88). The title compound 88 was prepared from

1-methoxy-3-vinylbenzene according to the same proce-

dure as described for the preparation of 86 from

1-methoxy-3-vinylbenzene. Compound 88 was then

used for the next reaction without further puriﬁcation:

1

white powder; TLC R_f=0.27 (n-hexane/EtOAc 1/3); H

NMR (300 MHz, CDCl₃) d 7.31–7.26 (m, 1H), 6.89–

6.82 (m, 3H), 6.07 (d, J=6.3 Hz, 1H), 5.7–5.01 (m, 1H),

3.91–3.85 (m, 2H), 3.80 (s, 3H), 2.65 (t, J=6.6 Hz, 1H),

2.24 (t, J=7.8 Hz, 2H), 1.67–1.58 (m, 2H), 1.29–1.27

(m, 8H), 0.87 (t, J=6.6 Hz, 3H); ee determination: the

corresponding 4-nitro benzoate prepared from 87 was

analyzed by HPLC: DAICEL CHIRALCEL OD

(0.46ꢄ25 cm); column temperature, 25 ^ꢁC; eluent,

EtOH/n-hexane=30/70; ﬂow rate, 0.5 mL/min; UV 260

nm; the retention times of 4-nitro benzoate and its

enantiomer were 11.7 and 19.7 min, respectively.

successivelywashed with H O and brine before being

2

dried over Na₂SO₄. Removal of the solvent byeva-

poration gave a residue, which was puriﬁed bycolumn

(2S)-2-(3-Methoxyphenyl)-2-octanoylaminoethyl disodium

phosphate (53). The title compound 53 was prepared

from 88, which was obtained from 1-methoxy-3-vinyl-

benzene, according to the same procedure as described

for the preparation of 1 from 60e (general method A):

beige powder, TLC R_f=0.26 (CHCl₃/MeOH/H₂O 65/

35/8); IR (KBr) 3390, 2929, 2857, 2213, 1637, 1547,

1491, 1466, 1437, 1262, 1092 cm^ꢀ1; ¹H NMR (300 MHz,

chromatographyon silica gel (Merck 7734, CHCl

/

3

MeOH/NH₄OH, 65/25/2-CHCl₃/MeOH/H₂O, 65/25/4)

to aﬀord a solid. The solid was dissolved in 1 M HCl/

EtOAc. After the organic layer was successively washed

with H₂O and brine, it was dried over Na₂SO₄.

Removal of the solvent byevaporation gave (2 S)-2-

octanoylamino-2-phenylethyl dihydrogen phosphate

(1.7 g, 68% yield from 90) which was converted to the

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

3785

disodium salt 52: white powder; TLC R_f=0.33 (CHCl₃/

MeOH/H₂O, 65/25/4); IR (KBr) 3368, 2923, 1645, 1562,

(300 MHz, CD₃OD) d 8.42 (brd, J=5.1 Hz, 1H), 7.73

(ddd, J=7.5, 7.5, 1.5 Hz, 1H), 7.47 (brd, J=7.8 Hz,

1H), 7.23 (ddd, J=7.5, 5.1, 1.2 Hz, 1H), 4.94 (dd,

J=6.3, 3.6 Hz, 1H), 4.17 (ddd, J=10.8, 6.6, 3.6 Hz,

1H), 4.08 (ddd, J=10.8, 7.2, 6.3 Hz, 1H), 2.40–2.24 (m,

2H), 1.61 (m, 2H), 1.40–1.20 (m, 8H), 0.89 (brt, J=6.6

Hz, 3H); MS (FAB, Pos.) m/z 411 (M+Na)⁺, 389

(M+H)⁺; HRMS (MALDI-TOF, Pos.) calcd for

1

1467, 1048, 963 cm^ꢀ1; H NMR (300 MHz, CD₃OD) d

7.40–7.20 (m, 5H), 5.19 (dd, J=7.5, 5.7 Hz, 1H), 4.20–

4.05 (m, 2H), 2.26 (t, J=7.5 Hz, 2H), 1.61 (quint,

J=7.5 Hz, 2H), 1.40–1.20 (m, 8H), 0.89 (t, J=6.6 Hz,

3H); [a]²_D⁵+41.2 (c1.1, MeOH); MS (FAB, Pos.) m/z 366

(M+Na)⁺, 344 (M+H)⁺, 246; HRMS (MALDI-TOF,

Pos.) calcd for C₁₆H₂₆NO₅P+Na⁺: 366.1446; found;

366.1401.

+

.

C₁₅H₂₃N₂O₅P 2Na+H : 389.1218; found; 389.1245.

2-Octanoylamino-2-(pyridin-3-yl)ethyl disodium phos-

phate (59). The title compound 59 was prepared from

92b according to essentiallythe same procedure as

described for the preparation on 1 from 60e (general

method A): 48% yield; colorless powder; TLC R_f=0.11

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3280, 2927,

N-(2-Hydroxy-1-pyridin-2-ylethyl)octanamide (92a). To

a stirred mixture of 91a²⁵(3.28 g, 10 mmol) and

N-methylmorpholine (3.3 mL, 30 mmol) in CH₂Cl₂(30

mL) was added dropwise octanoyl chloride (1.62 g, 10

mmol) at 0 ^ꢁC and stirring was continued at that tem-

perature for 2 h. The reaction mixture was quenched

with 1 M NaOH and extracted with CH₂Cl₂(3 times).

The combined organic layers were dried over MgSO₄.

Removal of the solvent byevaporation aﬀorded a resi-

due, which was puriﬁed bycolumn chromatographyon

silica gel (Merck 7734, n-hexane/EtOAc, 3/2–1/1–1/2–1/

3) to aﬀord ethyl 2-octanonylamino-2-(pyridin-2-yl)ace-

tate (1.16 g, 38% in 2 steps): TLC R_f=0.25 (n-hexane/

1650, 1547, 1467, 1093 cm^ꢀ^{1 1}H NMR (200 MHz,

;

CD₃OD) d 8.54 (d, J=1.6 Hz, 1H), 8.36 (dd, J=5.0, 1.6

Hz, 1H), 7.88 (brd, J=8.2 Hz, 1H), 7.36 (dd, J=8.2, 5.0

Hz, 1H), 4.93 (dd, J=6.4, 3.8 Hz, 1H), 4.18–3.89 (m,

2H), 2.33–2.24 (m, 2H), 2.40–2.24 (m, 2H), 1.70–1.50

(m, 2H), 1.40–1.15 (m, 8H), 0.87 (brt, J=6.6 Hz, 3H);

MS (FAB, Pos.) m/z 389 (M+H)⁺; HRMS (MALDI-

TOF, Pos.) calcd for C₁₅H₂₃N₂O₅P^ꢃ2Na+H⁺:

389.1218; found; 389.1220.

1

EtOAc, 2/1); H NMR (300 MHz, CDCl₃) d 8.55 (ddd,

J=4.8, 1.8, 0.9 Hz, 1H), 7.73 (ddd, J=7.5, 7.5, 1. 8 Hz,

1H), 7.50 (brd, J=7.5 Hz, 1H), 7.28–7.24 (m, 2H), 5.68

(d, J=6.6 Hz, 1H), 4.25–4.10 (m, 2H), 2.33–2.28 (m,

2H), 1.70–1.60 (m, 2H), 1.40–1.20 (m, 8H), 1.23 (t,

J=7.2 Hz, 3H), 0.86 (brt, J=6.6 Hz, 3H). The title

compound 92a was prepared from 2-octanonylamino-2-

(pyridin-2-yl)ethyl acetate according to the same proce-

dure as described for the preparation of 64a from

methyl (2-methoxyphenyl)(octanoylamino)acetate: 46%

Biological assay method

Inhibition of LPS-induced plasma TNF-ꢀ production in

rats. LPS from Escherichia coli strain 055 B5 (Difco

laboratories) and the test compounds were dissolved in

saline. Male Sprague–Dawley(CD)/IGS rats (Charles

River Inc., Japan) aged 6–8 weeks (n=5) were injected

intravenouslywith the test compounds (0.01–0.1 mg/ 10

mL/kg), and then immediatelygiven an intraperitoneal

injection of LPS (30 mg/kg). Plasma TNF-a production

was determined 90 min after the LPS challenge by

ELISA using a commercial kit (R&D Systems). ID₅₀

values were determined bylog-linear regression analysis

(3–4 doses per compound). ID₅₀=The dosage required

to inhibit plasma TNF-a production by50%. The data

were expressed as the meanꢂSEM of 5 animals per

group or ID₅₀values.

1

yield, TLC R_f=0.18 (n-hexane/EtOAc, 1/2); H NMR

(200 MHz, CDCl₃) d 8.51 (ddd, J=5.2, 1.8, 1.0 Hz, 1H),

7.72 (ddd, J=7.6, 7.6, 1.8 Hz, 1H), 7.40 (ddd, J=7.6,

1.0, 1.0 Hz, 1H), 7.25 (ddd, J=7.6, 5.2, 1.0 Hz, 1H),

7.07 (brd, J=7.0 Hz, 1H), 5.16 (ddd, J=7.0, 4.2, 4.2

Hz, 1H), 4.04 (dd, J=11.0, 4.2 Hz, 1H), 3.88 (dd,

J=11.0, 4.2 Hz, 1H), 3.20 (brs, 1H), 2.26 (dd, J=8.4,

6.2 Hz, 2H), 1.64 (m, 2H), 1.40–1.10 (m, 8H), 0.86 (brt,

J=6.6 Hz, 3H).

N-(2-Hydroxy-1-pyridin-3-ylethyl)octanamide (92b). The

title compound 92b was prepared from 91b²⁶according

to the same procedure as described for the preparation

of 92a from 91a: 42% yield; TLC R_f=0.38 (n-hexane/

% Inhibition=100- (C-S)/(L-S)ꢄ100.

C: Plasma TNF-a concentration of LPS-treated

animals pretreated with a test compound.

L: Plasma TNF-a concentration of LPS-treated

animals pretreated with saline.

1

EtOAc, 1/2); H NMR (300 MHz, CDCl₃) ꢀ 8.63 (brd,

J=1.8 Hz, 1H), 8.56 (dd, J=4.8, 1.8 Hz, 1H), 7.69

(ddd, J=8.1, 1.8, 1.8 Hz, 1H), 7.29 (dd, J=8.1, 4.8 Hz,

1H), 6.65 (brd, J=6.6 Hz, 1H), 5.62 (d, J=6.6 Hz, 1H),

4.30–4.15 (m, 2H), 2.26 (brt, J=8.1 Hz, 2H), 1.70–1.60

(m, 2H), 1.40–1.20 (m, 8H), 1.11 (t, J=7.2 Hz, 3H), 0.86

(brt, J=7.2 Hz, 3H).

S: Plasma TNF-a concentration of saline-treated

animals also pretreated with saline.

Inhibition of LPS-induced plasma TNF-ꢀ production in

mice. Male BALB/c mice aged 8 weeks (n=5) were

injected intravenouslywith the test compounds (0.01–

0.1 mg/ 10 mL/kg), and then immediatelygiven an

intraperitoneal injection of LPS (5 mg/10 mL/kg).

Plasma TNF-a production was determined 90 min after

the LPS challenge byELISA using a commercial kit

(GENZYME). ID₅₀values were determined bylog-linear

regression analysis (3–4 doses per compound).

2-Octanoylamino-2-(pyridin-2-yl)ethyl disodium phos-

phate (58). The title compound 58 was prepared from

92a according to essentiallythe same procedure as

described for the preparation on 1 from 60e (general

method A). 64% yield; colorless powder; TLC R_f=0.12

(CHCl₃/MeOH/H₂O, 65/25/4); IR (KBr) 3282, 2928,

1650, 1596, 1547, 1469, 1094, 985 cm^ꢀ^{1 1}H NMR

;

3786

T. Matsui et al. / Bioorg. Med. Chem. 10 (2002) 3757–3786

ID₅₀=the dosage required to inhibit plasma TNF-a

production by50%. The data were expressed as the

meanꢂSEM of 5 animals per group or ID₅₀values.

11. Nelson, F. C.; Zask, A. Exp. Opin. Invest. Drugs 1999, 8,

383.

12. Lowe, C. Exp. Opin. Ther. Patents. 1998, 8, 1309.

13. Shire, M. G.; Muller, G. W. Exp. Opin. Ther. Patents

1998, 8, 531.

14. Matsui, T.; Kondo, T.; Nishita, Y.; Itadani, S.; Nakatani,

S.; Omawari, N.; Sakai, M.; Nakazawa, S.; Ogata, A.; Ohno,

H.; Obata, T.; Nakai, H.; Toda, M. Bioorg. Med. Chem. Lett.

2002, 12, 903.

15. Matsui, T.; Kondo, T.; Nishita, Y.; Itadani, S.; Tsuruta,

H.; Fujita, S.; Omawari, N.; Sakai, M.; Nakazawa, S.; Ogata,

A.; Mori, H.; Ohno, H.; Obata, T.; Nakai, H.; Toda, M.

Bioorg. Med. Chem. Lett. 2002, 12, 907.

% Inhibition=100- (C-S)/(L-S)ꢄ100

C: Plasma TNF-a concentration of LPS-treated

animals pretreated with a test compound.

L: Plasma TNF-a concentration of LPS-treated

animals pretreated with saline

S: Plasma TNF-a concentration of saline-treated

animals also pretreated with saline.

16. Steiger, R. E. Org. Synth. 1995, III, 84.

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18. Dolle, R. E.; Schmidt, S. J.; Kruse, L. I. J. Chem. Soc.,

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Evaluation of TNF-ꢀ expression in mouse liver and

spleen by RT-PCR.³⁸The expression of TNF-a and

b-actin in the liver and spleen of BALB/c female mice

was evaluated bythe RT-PCR method. BALB/c mice

were obtained from Charles River (Shizuoka, Japan).

Mouse TNF-a and b-actin primers (Nihon bio-service,

Asaka City, Saitama, Japan) were used as speciﬁed by the

vendor. After 90 min of LPS (from W E. Coli O55:B5,

DIFCO LABORATORIES, Detroit, MI, USA) and drug

administration, each animal was decapitated. PGE₂was

used as the positive control (1 mg/kg, ip, 1 min before LPS

injection). Their liver and spleen were then removed,

rinsed in sterile saline, and immediatelystored at ꢀ80^ꢁC.

Total RNA was extracted with RNAzol B (TEL-TEST,

INC., Friendswood, TX, USA) according to the manu-

facturer’s instructions. Reverse transcription followed by

32 cycles (94^ꢁC for 1 min, 55^ꢁC for 1 min and 72^ꢁC for 3

min) of PCR were carried out with the ﬁrst-strand cDNA

synthesis kit (TaKaRa, Otsu, Shiga, Japan), the TaKaRa

Taq kit (TaKaRa, Otsu, Shiga, Japan) and a GeneAmp

PCR System 9600 thermal controller (PERKIN ELMER,

Foster City, Calif., USA), according to the manufac-

turer’s instructions. The PCR products were electro-

phoresed on a 1.5% agarose gel, stained with a 0.5 mg/mL

solution of ethidium bromide, and photographed under

UV transillumination.

19. Bannwarth, W.; Trzeciak, A. Helvetica Chimica Acta

1987, 70, 175.

20. Lee, V.; Newman, M. S. Org. Synth. 1988, VI, 613.

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˚

22. Wolfe, J. P.; Ahman, J.; Sadighi, J. P.; Singer, R. A.;

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28. 2,2-Dimethyloctanoyl chloride was prepared from

2-methylpropanoic acid by following successive procedures: (i)

LDA, C₆H₁₃I, (ii) SOCl₂.

29. 2-Trimethyleneoctanoyl chloride was prepared from

cyclobutanecarboxylic acid by following successive proce-

dures: (i) LDA, C₆H₁₃I, (ii) SOCl₂.

30. 2-Methyloctanoyl chloride was prepared from propionic

acid byfollowing successive procedures: (i) LDA, C H₁₃I, (ii)

6

SOCl₂.

31. (Pentyloxy)acetic acid was prepared from ethylene glycol

.

byfollowing successive procedures: (i) DHP, TsOH H₂O, (ii)

References and Notes

1. Tracey, K. J.; Cerami, A. Annu. Rev. Med. (45), 491.

2. Bemelmans, M. H.; van Tits, L. J.; Buurman, W. A. Crit.

Rev. Immunol. 1996, 16, 1.

NaH, C₅H₁₁I, (iii) TsOH, MeOH, (iv) Jone’s oxidation.

32. 4-Propoxybutanoic acid was prepared from butane-1,4-

.

diol byfollowing successive procedures: (i) DHP, TsOH H₂O,

3. Aggarwal, B. B.; Kohr, W. J.; Hass, P. E.; Moﬀat, B.;

Spencer, S. A.; Henzel, W. J.; Bringman, T. S.; Nedwin, G. E.;

Goeddel, D. V.; Harkins, R. N. J. Biol. Chem. 1985, 260, 2345.

4. Camussi, G.; Lupia, E. Drugs 1998, 55, 613.

5. Tilg, H. Can. J. Gastroenterol. 2001, 15, 661.

6. Jacobs, R. F.; Tabor, D. R. Scand. J. Infect. Dis. Suppl.

1990, 73, 7.

(ii) NaH, C₃H₇Br, (iii) TsOH, MeOH, (iv) Jone’s oxidation.

33. 5-Ethoxypentanoic acid was prepared from pentane-1,5-

.

diol byfollowing successive procedures: (i) DHP, TsOH H₂O,

(ii) NaH, C₂H₅Br, (iii) TsOH, MeOH, (iv) Jone’s oxidation.

34. 6-Methoxyhexanoic acid from hexane-1,6-diol by follow-

.

ing successive procedures: (i) DHP, TsOH H₂O, (ii) NaH,

C₂H₅Br, (iii) TsOH, MeOH, (iv) Jone’s oxidation.

7. Arrieta, O.; Rodriguez-Reyna, T. S.; Sotelo, J. Exp. Opin.

Ther. Patents 2000, 10, 601.

8. Blam, M. E.; Stein, R. B.; Lichtenstein, G. R. Am. J. Gas-

troenteral. 2001, 96, 1977.

35. Dibenzylphosphorochloridate was prepared from benzyl

phenoxymethylphosphinate according to the following paper

and used as 1 M stock solution: Kenner, G. W.; Todd, A. R.;

Weymouth, F. J. J. Chem. Soc. 1952, 3675.

9. Assche, V. G.; Rutgeerts, P. Exp. Opin. Invest. Drugs 2000,

9, 103.

36. Khorana, H. G.; Todd, A. R. J. Chem. Soc. 1953, 2257.

37. Gajda, T.; Zwierzak, A. Synthesis 1976, 243.

10. Marriott, J. B.; Westry, M.; Dalgleish, A. G. Drug Disc.

Today 1997, 2, 273.

38. David, S. A.; Silverstein, R.; Amura, C. R.; Kielian, T.;

Morrison, D. C. Antimicrob. Agents Chemother. 1999, 43, 912.

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