55399-93-4

Basic information

• Product Name: 4-HYDROXYISOLEUCINE
• Synonyms: HIL;HYDROXYISOLEUCINE, 4-;2-AMINO-2,3,5-TRIDEOXY-3-METHYL-D-XYLONIC ACID;(2S,3R,4S)-4-HYDROXYISOLEUCINE MAJOR ISOMER AND (2R,3R,4S)-4-HYDROXYISOLEUCINE MINOR ISOMER;[DISCONTINUED] Replaced by KIT-00008502-0HK;HYDROXYISOLEUCINE, 4-(FG);(2S,3R,4S)-2-Amino-4-hydroxy-3-methylpentanoic acid;(4S)-4-Hydroxy-L-isoleucine
• CAS NO: 55399-93-4
• Molecular Formula: C₆H₁₃NO₃
• Molecular Weight: 147.17
• EINECS: 2017-001-1
• Product Categories: chemical reagent;pharmaceutical intermediate;phytochemical;reference standards from Chinese medicinal herbs (TCM).;standardized herbal extract
• Mol File: 55399-93-4.mol

Chemical Properties

• Melting Point: 223-224℃
• Boiling Point: 331.6ºCat 760 mmHg
• Flash Point: 147℃
• Appearance: /
• Density: 1.181±0.06 g/cm3 (20 ºC 760 Torr)
• Vapor Pressure: 1.11E-05mmHg at 25°C
• Refractive Index: 1.495
• Storage Temp.: 2-8°C
• Solubility: Methanol (Slightly), Water (Slightly)
• PKA: 2.41±0.25(Predicted)
• BRN: 4128096
• CAS DataBase Reference: 4-HYDROXYISOLEUCINE(CAS DataBase Reference)
• NIST Chemistry Reference: 4-HYDROXYISOLEUCINE(55399-93-4)
• EPA Substance Registry System: 4-HYDROXYISOLEUCINE(55399-93-4)

Safety Data

• Hazard Codes: Xi
• Statements: 36/37/38
• Safety Statements: 26
• WGK Germany: 3
• Hazardous Substances Data: 55399-93-4(Hazardous Substances Data)

Usage

Uses

Used in Pharmaceutical Industry:
4-Hydroxyisoleucine is used as a therapeutic agent for the treatment of type II diabetes. It facilitates an increase in glucose-stimulated insulin release from isolated rat islets and perfused rat pancreas at a concentration of 200 μM. Additionally, it enhances PI3K activity in muscle and liver tissues in rats, particularly in a rat model of type 2 diabetes induced by nicotinamide and streptozotocin (STZ) when administered at 25 mg/kg. Furthermore, it effectively decreases plasma glucose, triglyceride, LDL, HDL, and cholesterol levels in an STZ-induced rat model of type 1 diabetes when administered at a dose of 50 mg/kg per day for four weeks.

InChI:InChI=1/C6H13NO3/c1-3(4(2)8)5(7)6(9)10/h3-5,8H,7H2,1-2H3,(H,9,10)

Upstream product

• 168610-73-9 (3SR,4RS,5SR)-3-amino-4,5-dimethyldihydrofuran-2-one 
• 885054-30-8 ethyl (2S,3R)-2-amino-3-methyl-4-oxopentanoate 
• 73-32-5 L-isoleucine 
• 950204-60-1 (2R,4S,5R,6S)-5,6-dimethyl-2-phenyl-tetrahydro-2H-1,3-oxazin-4-carboxylic acid 
• 950204-59-8 (2R,4S,5R,6S)-2-cyclohexyl-5,6-dimethyl-tetrahydro-2H-1,3-oxazin-4-carboxylic acid 
• 950204-27-0 (2R,4S,5R,6S)-5,6-dimethyl-2-(4-nitrophenyl)-tetrahydro-2H-1,3-oxazin-4-carboxylic acid 
• 942208-47-1 C₇H₁₂N₂O₃ 
• 55527-86-1 (3S,4R,5S)-3-amino-4,5-dimethyldihydrofuran-2(3H)-one 
• 55527-85-0 (3S,4R,5R)-3-Amino-4,5-dimethyl-dihydro-furan-2-one 
• 168610-73-9 4-hydroxyisoleucine γ-lactone 
• 600-18-0 2-Oxobutyric acid 
• 75-07-0 acetaldehyde 
• 21704-91-6 (3S,4R,5S)-3-amino-4,5-dimethyl-3,4-dihydro-2(5H)-furanone hydrochloride 
• 1093653-23-6 C₇H₁₁NO₃ 

Downstream Products

• 950204-27-0 (2R,4S,5R,6S)-5,6-dimethyl-2-(4-nitrophenyl)-tetrahydro-2H-1,3-oxazin-4-carboxylic acid 
• 950204-60-1 (2R,4S,5R,6S)-5,6-dimethyl-2-phenyl-tetrahydro-2H-1,3-oxazin-4-carboxylic acid 
• 950204-59-8 (2R,4S,5R,6S)-2-cyclohexyl-5,6-dimethyl-tetrahydro-2H-1,3-oxazin-4-carboxylic acid 
• 908856-77-9 (3S,4R,5S)-3-(dibenzylamino)-4,5-dimethyldihydrofuran-2(3H)-one 
• 908856-76-8 C₂₇H₃₁NO₃ 
• 71392-28-4 (3S,4S,5R)-3-amino-4,5-dimethyldihydrofuran-2(3H)-one 
• 908856-93-9 C₁₄H₁₇NO₄ 

Relevant articles and documents

Engineering Bacillus subtilis Isoleucine Dioxygenase for Efficient Synthesis of (2 S,3 R,4 S)-4-Hydroxyisoleucine

Isoleucine dioxygenase (IDO)-catalyzed hydroxylation of isoleucine is a promising method for the synthesis of the diabetic drug (2S,3R,4S)-4-hydroxyisoleucine [(2S,3R,4S)-4-HIL]. However, the low activity of IDO significantly limits its practical application. In this work, a high-throughput screening method was developed and directed evolution was performed on the IDO from Bacillus subtilis, resulting in a double mutant with improvements in specific activity, protein expression level, and fermentation titer of 3.2-, 2.8-, and 9.4-fold, respectively. l-Isoleucine (228 mM) was completely converted to (2S,3R,4S)-4-HIL by the best variant with a space-time yield of up to 80.8 g L-1 d-1, which is the highest record reported so far. With a further increase of the substrate loading to 1 M, a high conversion of 91% could also be achieved. At last, enzymatic synthesis of (2S,3R,4S)-4-HIL was successfully carried out on a 3 L scale, indicating tremendous potential of the IDO variant I162T/T182N for green and efficient production of (2S,3R,4S)-4-HIL.

Repurposing Nonheme Iron Hydroxylases to Enable Catalytic Nitrile Installation through an Azido Group Assistance

Three mononuclear nonheme iron and 2-oxoglutarate dependent enzymes, l-Ile 4-hydroxylase, l-Leu 5-hydroxylase and polyoxin dihydroxylase, are previously reported to catalyze the hydroxylation of l-isoleucine, l-leucine, and l-α-amino-δ-carbamoylhydroxyvaleric acid (ACV). In this study, we showed that these enzymes can accommodate leucine isomers and catalyze regiospecific hydroxylation. On the basis of these results, as a proof-of-concept, we demonstrated that the outcome of the reaction can be redirected by installation of an assisting group within the substrate. Specifically, instead of canonical hydroxylation, these enzymes can catalyze non-native nitrile group installation when an azido group is introduced. The reaction is likely to proceed through C - H bond activation by an Fe(IV)-oxo species, followed by azido-directed C-N bond formation. These results offer a unique opportunity to investigate and expand the reaction repertoire of Fe/2OG enzymes.

Attempt to simultaneously generate three chiral centers in 4-hydroxyisoleucine with microbial carbonyl reductases

A panel of microorganisms was screened for selective reduction ability towards a racemic mixture of prochiral 2-amino-3-methyl-4-ketopentanoate (rac-AMKP). Several of the microorganisms tested produced greater than 0.5 mM 4-hydroxyisoleucine (HIL) from rac-AMKP, and the stereoselectivity of HIL formation was found to depend on the taxonomic category to which the microorganism belonged. The enzymes responsible for the AMKP-reducing activity, ApAR and FsAR, were identified from two of these microorganisms, Aureobasidium pullulans NBRC 4466 and Fusarium solani TG-2, respectively. Three AMKP reducing enzymes, ApAR, FsAR, and the previously reported BtHILDH, were reacted with rac-AMKP, and each enzyme selectively produced a specific composition of HIL stereoisomers. The enzymes appeared to have different characteristics in recognition of the stereostructure of the substrate AMKP and in control of the 4-hydroxyl group configuration in the HIL product.

A practical and efficient total synthesis of potent insulinotropic (2S,3R,4S)-4-Hydroxyisoleucine through a chiral N-protected γ-keto- α-aminoester

(2S,3R,4S)-4-Hydroxyisoleucine, which exhibits remarkable insulinotropic activity, is expected to be a potent drug to treat type II diabetes. We propose herein a four-step synthesis of the enantiopure natural product on the basis of successive Mannich condensation, catalytic epimerization, N-paramethoxyphenyl deprotection, and diastereoselective reduction. This compact economical and scalable sequence enables to perfectly control three contiguous chiral centers. It does not involve any chromatographic purification, and the desired compound is obtained in >99 % de, >99 % ee, and 22 % overall yield under our optimized conditions.

An organocatalyzed enantioselective synthesis of (2S,3R,4S)-4- hydroxyisoleucine and its stereoisomers

A concise enantioselective total synthesis of (2S,3R,4S)-4- hydroxyisoleucine and its stereoisomers is described. A key feature of this protocol is a catalytic enantioselective mannich reaction that is either anti- or syn-selective as genesis of chirality.

METHOD FOR PURIFYING 4-HYDROXYISOLEUCINE

The present invention aims to provide a method of conveniently isolating and purifying as well as separating and removing (2S,3R,4S)-4-hydroxyisoleucine at a high purity and in a high yield. Specifically, the present invention discloses a purification method of (2S,3R,4S)-4-hydroxyisoleucine or a chemically acceptable salt thereof, which includes the following steps (a), (b) and (c): (a) a step of reacting (2S,3R,4S)-4-hydroxyisoleucine or a chemically acceptable salt thereof in a mixture with an aldehyde compound represented by the formula: Q-CHO wherein Q is an aryl group having a carbon number of 6 to 14, an alkyl group having a carbon number of 1 to 10, a cycloalkyl group having a carbon number of 3 to 10 or a 5- to 10-membered heterocyclic group, each of which optionally has substituent(s), or an equivalent form thereof to give a compound represented by the formula (1) wherein Q is as defined above, (b) a step of extracting the compound represented by the formula (1) with an organic solvent, and (c) a step of converting the compound represented by the formula (1) to (2S,3R,4S)-4-hydroxyisoleucine or a chemically acceptable salt thereof.

Synthesis of hydantoin analogues of (2S,3R,4S)-4-hydroxyisoleucine with insulinotropic properties

The first synthesis of an optically pure (2R,3R,4S)-hydantoin 2, analogue of (2S,3R,4S)-4-hydroxyisoleucine, was achieved in two steps in un-optimized 35% overall yield from previously reported aldehyde synthon 1. (2R,3R,4S)-Hydantoin is stable at acidic pH. This solves the major drawback of (2S,3R,4S)-4-hydroxyisoleucine that easily cyclizes into inactive lactone. Furthermore, (2R,3R,4S)-hydantoin stimulates the insulin secretion by 150% at 25 μM compared with 4-hydroxyisoleucine and insulin secretagogue drug repaglinide. In view of its stability and biological activity, (2R,3R,4S)-hydantoin represents a good candidate for type-2 diabetes management and control.

METHODS FOR THE SYNTHESIS OF 4-HYDROXYISOLEUCINE, STEREOISOMERS AND ANALOGS THEREOF

A method for synthesizing 4-hydroxyisoleucine, stereoisomers and analogs thereof having a general formula I is disclosed herein. The method comprises reacting an alkyl isocyanoacetate of formula III with an acetoin or an acetoin analog of formula II to give an unsaturated N-formyl lactone of formula IV, hydrogenating the unsaturated N-formyl lactone to give N-formyl lactone of formula V and hydrolysing said lactone to yield the desired product of formula I.

Synthesis of 4-hydroxyisoleucine by the aldolase-transaminase coupling reaction and basic characterization of the aldolase from Arthrobacter simplex AKU 626

Arthrobacter simplex AKU 626 was found to synthesize 4-hydroxyisoleucine from acetaldehyde, α-ketobutyrate, and L-glutamate in the presence of Escherichia coli harboring the branched chain amino acid transaminase gene (ilvE) from E. coli K12 substrain MG-1655. By using resting cells of A. simplex AKU 626 and E. coli BL21(DE3)/pET-15b-ilvE, 3.2 mM 4-hydroxyisoleucine was produced from 250 mM acetaldehyde, 75 mM α-ketobutyrate, and 100mM L-glutamate with a molar yield to α-ketobutyrate of 4.3% in 50 mM Tris-HCl buffer (pH 7.5) containing 2 mM MnCl2·4H2O at 28°C for 2 h. An aldolase that catalyzes the aldol condensation of acetaldehyde and α-ketobutyrate was purified from A. simplex AKU 626. Mn2+ and pyridoxal 5′-monophosphate were effective in stabilizing the enzyme. The native and subunit molecular masses of the purified aldolase were about 180 and 32 kDa respectively. The N-terminal amino acid sequence of the purified enzyme showed no significant homology to known aldolases.

Method for the synthesis of 4-hydroxyisoleucine and the derivatives thereof

The invention relates to a method for the synthesis of two isomers, at function OH, alone or in mixtures, of amino acids α or the derivatives thereof, having general formula (B), wherein: linkage C—O of the 4-position carbon (represented by symbol) denotes one or other of isomers III or IV, or mixtures thereof. Moreover, R1 and R2 represent: a hydrogen atom; or either R1 or R2 represents a hydrogen atom and the other substituent is a radical Ra, an acyl group —CORa, such as acetyl, or a functional group —COORa, —SO2Ra, —N (Ra, Rb), Ra and Rb, which are identical or different, representing a C1-C12 linear or branched alkyl radical, optionally substituted, an aryl group with one or more aromatic rings and heterocycles, comprising between 5 and 8C, optionally substituted, or aralkyl, the alkyl substituent and the aryl group being as defined above; or R1 and R2 both represent a substituent as defined above. R3 represents a hydrogen atom or Ra and R4 has the significance of Ra. The invention is characterised in that it comprises: the isomerisation of a compound having formula (I), wherein R1, R2, Ra, R3 and R4 are as defined above, such as to produce a compound having formula (II); and the reduction of the carbonyl function thereof which, depending on the catalytic system employed and the formula (I) compound used, produces one of the isomers having general formula (III) or (IV) or a mixture thereof having formula (B). The invention can be used for the synthesis of (2S, 3R, 4S)-4-hydroxyisoleucine.