81093-37-0

Usage

New Hypolipidemic drugs

Pravastatin is a derivative of Vastatin. It is a new type of hypolipidemic drug. It is especially suitable for primary hypercholesterolemia which is still not controlled by dietary restriction. The drug is statins, developed by Japan Sankyo Pharmaceutical Co. Ltd. and AmericanBristo1-Myers Sguibb Company. It was listed in Japan for the first time in 1989. It is a new drug of Category 4 of western medicine in China, which has a broad market prospect.

Pharmacological action

Pravastatin is a cholesterol lowering drug and is a competitive inhibitor of the Hydroxymethyl glutaryl coenzyme A reductaseh (HMG-CoA reductase). The mechanism of action is the same with lovastatin. It can inhibit the synthesis of cholesterol, increase the metabolism of low density lipoprotein (LDL) and reduce the cholesterol. It can also increase the high density lipoprotein (HDL) and reduce triglyceride. The efficacy of this product is the same as lovastatin and statins. It is used for heterozygous familial, non familial hypercholesterolemia and mixed hyperlipidemia, familial hyperlipoproteinemia type III and serum hypertriglyceridemia (type IV). Pravastatin is an open - ring active substance, and its sodium salt is used clinically. Like other statins, this product competitively inhibits HMG-CoA reductase, reduces serum TC, LDL-C and ApoB, also decreases serum VLDL and TG, and increases HDLC and Apo-A, which is similar to lovastatin.

Pharmacokinetics

This product is absorbed orally, and its bioavailability is about 17%. The concentration of the blood drug peak is reached 1 to 1.5 hours after taking the medicine. It can be selectively distributed in liver cells. It is metabolized in the liver, the active metabolite is TK 2.5% ~ 10%. It excretes rapidly. After one oral administration, 20% and 71% is discharged through urine and feces within 96 hours. The half-life is 1.5 to 2 hours.

Adverse reaction

The occasional adverse reaction includes diarrhea, abdominal pain, and gastrointestinal discomfort as well as the rise of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase, gamma glutamyl transpeptidase. Other occasional adverse reaction includes the increase of CPK and urea nitrogen and the phenomenon of urine occult blood.

Precaution

The curative effect of homozygous family hypercholesterolemia is poor. During the treatment, the liver function should be checked regularly. If the increase of SGPT and SGOT is equal to or more than three times the normal upper limit and is persistent, the treatment should be stopped. Patients with the history of liver disease or drinking history should be careful in use. The use of HMG-COA reductase inhibitor class lipid lowering drugs can cause the increase of CPK. If the rise value is 10 times the normal limit, it should be stopped. In the course of use, if the patient has unexplained myalgia, pain, weakness, especially those with discomfort and fever, they should be reported to the doctor immediately. Other HMG-COA reductase inhibitors such as cyclosporin, fibrinic acid derivatives, nicotinic acid, etc. can increase the incidence of myositis and myopathy.However, if pravastatin is used simultaneously with the above drugs, clinical trials indicates that the incidence of myositis and myopathy will not be increased.

Originator

Sankyo (Japan)

Manufacturing Process

Pravastatin was isolated as products of enzymatic hydroxylation by some kinds of microorganisms of [1S-[1-α(R*),7β,8β(2S*,4S*)8αβ]]-2methylbutanoic acid 1,2,3,7,8,8a-hexahydro-7-methyl-8-[2-(tetrahydro-4hydroxy-6-oxo-2))-pyran-2-yl)ethyl]-1-naphthalenic lactone (campactin) or their carboxylic acid or their salts (products of animal metabolism of microorganisms from the genera Nocardia, Streptomyces et cetera).Pravastatin may be preparated by using the microorganisms of genera Nocardia (method 1) and Mortierella (method 2).Method 1 Cultivation of Nocardia autotrophica subsp. amethystineCells of Nocardia autotrophica subsp. amethystina FERM P-6183 was inoculated from a slant culture by means of a platinum loop into each of twenty 500 ml Erlenmeyer flasks, each containing 100 ml of a culture medium having the following composition: glucose - 1.0%, peptone - 0.2%, meat extract - 0.1%, yeast extract - 0.1%, corn steep liquor 0.3%, tap water balance..Shaking was then carried out at 26°C and 220 r.p.m. for 2 days, at which time sodium 2-methyl-8-(2-methyl-1-oxobutoxy)-β,δ-dihydroxy(1S-(1-α(βS*,δ-S*),2-α,6-α,8-β(R*),8a-α))-1-1,2,6,7,8,8ahexahydronaphthaleneheptanoate was added to a final concentration of 0.05% w/v. Incubation was continued at 26°C and 220 r.p.m. for a further 5 days.Preparation of pravastatinAfter completion of the cultivation, the reaction mixture was filtered and the pH of the filtrate was adjusted to a value of 3 by the addition of trifluoroacetic acid. The acidified filtrate was then extracted three times, each with 1 liter of ethyl acetate, to give extracts containing a mixture (6-α and 6-β) of (1S-(1α,β- S*,δ-S*),2-α,8-βR*),8a-α))-1-naphthaleneheptanoic acid 1,2,6,7,8,8ahexahydro-2-methyl-8-(2-methyl-1-oxobutoxy)-β,δ,6-trihydroxy.This extract was then immediately transferred into a 5% w/v aqueous solution of sodium hydrogen carbonate, and the pH of the mixture was adjusted to a value of 7.0 by the addition of 2 N hydrochloric acid. The mixture was then adsorbed on a Diaion HP-20 column. The column was washed with water and then eluted with 50% v/v aqueous acetone to give a fraction containing (1S(1-α,β- S*,δ-S*),2-α,6-α,8-βR*),8a-α))-1-naphthaleneheptanoic acid 1,2,6,7,8,8a-hexahydro-2-methyl-8-(2-methyl-1-oxobutoxy)-β,δ,6-trihydroxy-, monosodium salt (pravastatin). This was freeze-dried, to give 200 mg of pravastatine.Method 2 Cultivation of Mortierella maculata nov. spec. E-97 [NCAIM(P)F 001266]A spore suspension was prepared with 5 ml of a 0.9% sodium chloride solution obtained from a 7-10 day old, malt extract-yeast extract agar slant culture of Mortierella maculata nov. spec. E-97 [NCAIM(P)F 001266] strain able to 6-β-hydroxylate compactin and the suspension was used to inoculate 100 ml inoculum medium PI (glucose-50 g, soybean meal-20 g, in 1000 ml tap water) sterilized in a 500 ml Erlenmeyer flask.5 liters working volume a bioconversion culture medium is prepared (glucose20 g, glycerine-20 g, soybean meal-20 g, peptone-5 g, potassium dihydrogen phosphate-0.5 g, polypropylene glycol 2000-1 g, in 1000 ml tap water); the components of the culture medium are added corresponding to 5 liters. Then it was sterilized for 45 min at 121°C and seeded with 500 ml of the inoculum culture.Before sterilization the pH of the medium was adjusted to 7.0 value.The fermentation was carried out at 28°C, with a stirring rate of 400 rpm andwith an aeration rate from bottom direction 60 liters/hour for 4 days. At the 2nd day after the transfer the culture started to foam heavily, which can be decreased by the addition of further polypropylene glycol 2000. The pH reached 6.3-7.5 by the 4th day. The feeding of the sodium 2-methyl-8-(2methyl-1-oxobutoxy)-β,δ-dihydroxy(1S-(1-α(β- S*,δ-S*),2-α,6-α,8-β(R*),8aα))-1-1,2,6,7,8,8a-hexahydronaphthaleneheptanoate substrate is allowed to be started if the pH of the broth is above 6.3.Preparation of pravastatinAt the 4th day of the fermentation 2.5 g compactin substrate is added in sterile filtered aqueous solution. Calculated for the volume of the broth 0.51.0% glucose was added into the culture depending on the pH in the form of 50% solution sterilized at 121°C for 25 min in parallel with the substrate feeding. After 24 hours the compactin substrate is consumed from the culture (is detected by HPLC) and was converted to pravastatin. By lyophilization of the aqueous residue 1.3 g pravastatin was obtained. The chromatographically pure product was crystallized from a mixture of ethanol and ethyl acetate. Melting point: 170-173°C (decomp.).

Therapeutic Function

Antihyperlipidemic

InChI:InChI=1/C23H36O7/c1-4-13(2)23(29)30-20-11-17(25)9-15-6-5-14(3)19(22(15)20)8-7-16(24)10-18(26)12-21(27)28/h5-6,9,13-14,16-20,22,24-26H,4,7-8,10-12H2,1-3H3,(H,27,28)/t13-,14-,16?,17+,18+,19-,20-,22-/m0/s1

Upstream product

• 84173-31-9 3,5-bis-epi-compactin 
• 81131-70-6 pravastatin sodium salt 
• 73573-88-3 mevastatin 

Downstream Products

• 81131-70-6 Sodium; 3,5-dihydroxy-7-[(1S,2S,6S,8S,8aR)-6-hydroxy-2-methyl-8-((S)-2-methyl-butyryloxy)-1,2,6,7,8,8a-hexahydro-naphthalen-1-yl]-heptanoate 
• 81131-70-6 pravastatin sodium salt 
• 135716-16-4 (S)-2-Methyl-butyric acid (1S,3R,4aS,7S,8S,8aS)-8-(2-{(4R,6S)-6-[2-(tert-butyl-diphenyl-silanyloxy)-ethyl]-2,2-dimethyl-[1,3]dioxan-4-yl}-ethyl)-3-hydroxy-7-methyl-decahydro-naphthalen-1-yl ester 
• 136491-73-1 (S)-2-Methyl-butyric acid (1S,4aS,7S,8S,8aS)-8-(2-{(4R,6S)-6-[2-(tert-butyl-diphenyl-silanyloxy)-ethyl]-2,2-dimethyl-[1,3]dioxan-4-yl}-ethyl)-7-methyl-3-oxo-decahydro-naphthalen-1-yl ester 
• 135716-15-3 (S)-2-Methyl-butyric acid (1S,3R,4aS,7S,8S,8aS)-8-[(3R,5S)-7-(tert-butyl-diphenyl-silanyloxy)-3,5-dihydroxy-heptyl]-3-hydroxy-7-methyl-decahydro-naphthalen-1-yl ester 
• 85956-22-5 pravastatin lactone 

Relevant academic research and scientific papers

He the sandbank contains the fluorine derivative and use thereof

The present invention belongs to the field of pharmaceutical chemistry, and provides 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor, wherein a fragment containing 3-fluoro-caprolactone and a lactone thereof are subjected to ring opening to form a poly-substituted pyrimidine statin fluorine-containing modifier of 1-fluoro-3-hydroxy-pentanoic acid and a salt or ester thereof, ie., the 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor, and the structure formula is defined in the specification. According to the present invention, the test results show that the compounds have the HMG-CoA reductase activity inhibition effects, and can be used as the new generation of the potential HMG-CoA reductase inhibitors.

Fermentation Medias and Processes Thereof

The present invention demonstrates the utility of carbonic acid amides such as urea or its derivatives, carbamates, carbodiimides & thiocarbamides as nitrogenous supplements in fermentation media for production of recombinant proteins to achieve enhanced bioconversion rates and peptides like insulin and insulin analogues, exendin and enzymes such as lipase using methanol inducible fungal expression systems such as Pichia.

In situ bioconversion of compactin to pravastatin by Actinomadura species in fermentation broth of Penicillium citrinum

The biocatalytic production of pravastatin from compactin by hydroxylation has found many applications in health care and pharmaceuticals. Actinomadura macra, Actinomadura madurae, and Actinomadura livida can efficiently bioconvert compactin to pravastatin. The fermentation broth (Penicillium citrinum fermented media) harvested on the eighth day contained 388.90 mg L-1 of compactin and an undetectable level of mycotoxin (citrinin). Bioconversion by A. macra was highest (87 %) in the yeast extract-amended medium. The anti-actinomadura effects of citrinin reduce the bioconversion capacity of Actinomadura. The in situ hydroxylation of compactin produced by P. citrinum represents a preferable alternative for the use of purified compactin, as a way to reduce cost and time processing.

Methods for predicting the response to statins

The invention provides methods for optimizing therapeutic efficacy for treating hypercholesterolemia in a subject having a cardiovascular disease (CVD), comprising (a) determining subject characteristics that affect the likelihood of reaching a goal level of low density lipoprotein (LDL); and (b) obtaining success probabilities of a variety of statin treatments for reaching said goal level of LDL using said subject characteristics and a multivariate model; and (c) administrating the optimal statin treatment with the highest success probability of step (b) to said subject thereby optimizing therapeutic efficacy for treating hypercholesterolemia in said subject.

Drug or Supplement Combination with Conjugated Linoleic Acid for Fat Loss in Mammals

Food, feed or drug combinations with conjugated linoleic acid are described that cause enhanced fat loss in mammals more efficiently than any of the individual components of the combination. Food, feed, or drugs that activate AMP activated protein kinase, agonists of nuclear receptors that bind RXR in adipocytes, or statin inhibitors were found to be more effective for fat loss when combined with conjugated linoleic acid.

Efficient biotransformations using Escherichia coli with tolC acrAB mutations expressing cytochrome P450 genes

We report here some efficient biotransformations using Escherichia coli strains with disruptions for the AcrAB-TolC efflux pump system. Biotransformations of compactin into pravastatin (6α-hydroxy-iso- compactin) were performed using E. coli strains with tolC and/or acrAB mutations expressing a cytochrome P450 (P450) gene. The production levels of pravastatin using strains with acrAB, tolC, and tolC acrAB mutations increased by 3.7-, 7.0-, and 7.1-fold, respectively. Likewise, the production levels of 25-hydroxy vitamin D3 and 25-hydroxy 4-cholesten 3-one using tolC acrAB mutant strains expressing an individual P450 gene increased by 2.2- and 16-fold, respectively. The enhancement of this biotransformation efficiency could be explained by increases in the intracellular amounts of substrates and the concentrations of active P450s. These results demonstrate that we have achieved versatile methods for efficient biotransformations using E. coli strains with tolC acrAB mutations expressing P450 genes.

Capillary electrophoresis determination of pravastatin and separation of its degradation products

A capillary zone electrophoresis method for pravastatin determination was developed and validated. Rapid migration of negatively charged pravastatin molecule was obtained in alkaline buffer by the application of electric field of 30 kV. Influence of the pH value and ionic strength of running buffer, applied voltage and capillary temperature on mobility and sensitivity was evaluated. Detection wavelength was set to 237 nm. The method was applied to the determination of the drug in pharmaceutical dosage form. Pravastatin is a δ-hydroxy acid, which is prone to lactonize and epimerize in a pH-dependent manner. Micellar electrokinetic chromatographic approach was chosen to develop a method able to separate pravastatin and its degradation products in acidic media. The proposed method allows baseline separation of hydroxy acid and neutral lactone forms of the drug that appear as interconversion products depending on the pH value.

PROCESS FOR THE PREPARATION OF PRAVASTATIN

The invention relates to a process for the preparation of pure pravastatin or a salt thereof using an adsorption chromatography technique. The invention also relates to pharmaceutical compositions that include the pure pravastatin and to use of the compositions for treating hypercholesterolemia.

Strains of saccharaothrix, process for producing pravastain using the strains and isolation process of (HMG)-CoA reductase

The present invention provides two new microorganism strains of Saccharothrix, designated as YS-44442 and YS-45494, a process of producing pravastatin using the strains, and an improved process for isolation of (HMG)-CoA reductase inhibitors.

Method of purifying pravastatin

The present invention provides pure pravastatin compositions and pure compactin compositions, and methods for the preparation thereof.