WO1989007607A1 - 24r-scymnol, and preparation and use thereof - Google Patents

24r-scymnol, and preparation and use thereof Download PDF

Info

Publication number
WO1989007607A1
WO1989007607A1 PCT/AU1989/000064 AU8900064W WO8907607A1 WO 1989007607 A1 WO1989007607 A1 WO 1989007607A1 AU 8900064 W AU8900064 W AU 8900064W WO 8907607 A1 WO8907607 A1 WO 8907607A1
Authority
WO
WIPO (PCT)
Prior art keywords
scymnol
preparation
activity
compound
hydrolysis
Prior art date
Application number
PCT/AU1989/000064
Other languages
French (fr)
Inventor
Yoshiki Kosuge
Takuo Kosuge
Kuniro Tsuji
Hitoshi Ishida
Original Assignee
Broadbent, James, Meredyth
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Broadbent, James, Meredyth filed Critical Broadbent, James, Meredyth
Priority to DE68925355T priority Critical patent/DE68925355T2/en
Priority to EP89902728A priority patent/EP0401262B1/en
Publication of WO1989007607A1 publication Critical patent/WO1989007607A1/en
Priority to DK192890A priority patent/DK167220B1/en
Priority to US08/176,576 priority patent/US5451575A/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics

Definitions

  • This invention relates to the compound 24R-scymnol, to the preparation of this compound in substantially pure form, and to the use thereof, for example in the treatment of liver dysfunction.
  • prior International Patent Application No. PCT/AU87/00281 there is disclosed a process for the isolation and preparation of an active principle by extraction from particular tissues of sharks.
  • scymnol is the mixture of 24R and 24S compounds, since theoretically the synthesized scymnol should be produced in both 24R and 24S configurations on reduction with NaBH 4 of the ketone in the 24-position of the starting material as described by Bridgwater et.al.
  • the present invention provides a method for the preparation of the compound of general formula I, which comprises the step of hydrolysis of a 24R-scymnol sulphate ester with an inorganic acid, preferably in the presence of barium chloride.
  • the process for the preparation of 24R-scymnol as broadly outlined above is particularly advantageous as it can be carried out using aqueous acids.
  • 24R-scymnol has been prepared by hydrolysis of sodium scymnol sulphate with dilute hydrochloric acid in the presence of barium chloride, to give a crystalline product.
  • Table 1 The physical and chemical data of the product are set out in Table 1:
  • bioassay measures tyrosine aminotransferase (TA) activity in liver of mice.
  • TA is one of important enzymes in liver.
  • ASSAY Mice (5 weeks old) were orally administered the active principle of shark's bile (MD) (lOmg/kg), 24R-scymnol (lOmg/kg), or water (6:00 PM on the previous day, 9:00 AM on the day). After one hour from the last administration, the mice were forced to swim in a water at 35°C. After 4 hours swimming, mice were sacrificed by decapitation and livers were perfused with 0.145 M KCl to remove blood. Half gram of liver was homogenized in 0.145 M KCl and centrifuged at 10,000xg for 30min. and TA activity in the supernatant was measured by the method of Diammondstone. The activity is shown as the amount of p-hydroxyphenyl pyruvic acid (p-HPP) produced by the enzyme reaction for lOmin.
  • p-HPP p-hydroxyphenyl pyruvic acid
  • the present invention also provides a pharmaceutical composition comprising 24R-scyir.no1, together with a pharmaceutically acceptable carrier or diluent therefor.
  • 24R-scymnol can be formulated as stable tablets after being mixed as a powder with a known carrier or bulking agent.
  • Such pharmaceutical compositions may be used, for example, for the activation of liver function in the treatment of the diseases of the liver such as hepatitis, nephritis, diabetes, etc. Further details of this invention will be apparent from the following Examples which illustrate the invention without limiting it in any way.
  • EXAMPLE 1 Preparation of 24R-scymnol.
  • the active principle of shark's bile (500mg) was dissolved in 7ml of 1% HC1 containing 400mg of BaCl 2 and the mixture was heated for 5h at 100°C. After cooling, the resulting solution was extracted three times with 50ml of AcOH-BuOH (1:1). The organic layer was washed twice with H 2 0. Removal of the solvent gave a yellow oil. The resultant residue was dissolved in MeOH and applied to reversed phase HPLC. 50mg of 24R-scymnol was obtained.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Animal Behavior & Ethology (AREA)
  • Engineering & Computer Science (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Veterinary Medicine (AREA)
  • Steroid Compounds (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Saccharide Compounds (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Polymerisation Methods In General (AREA)
  • Glass Compositions (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Compounds Of Unknown Constitution (AREA)
  • Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Inks, Pencil-Leads, Or Crayons (AREA)
  • Luminescent Compositions (AREA)

Abstract

24R-Scymnol, having general formula (I).

Description

24R-SCYMNO , AND PREPARATION AND USE THEREOF
This invention relates to the compound 24R-scymnol, to the preparation of this compound in substantially pure form, and to the use thereof, for example in the treatment of liver dysfunction. in prior International Patent Application No. PCT/AU87/00281 there is disclosed a process for the isolation and preparation of an active principle by extraction from particular tissues of sharks. This active principle, now termed "isolutrol", was isolated in good yield from an aqueous extract of the livers and/or gall bladders of sharks, and the active component therein identified as 2_R-(+)-3α,7α,12α,24,26- penta- hydroxycoprostane-27-sodium sulphate ester (sodium 24R-scymnol sulphate) . It has now been found that 24R-scymnol can be prepared from the active component disclosed in the prior International Patent Application, and that 24R-scymnol has activity in the treatment of liver dysfunction. According to a first aspect of the present invention, there is provided the compound of the general formula I, in substantially pure form:
Figure imgf000004_0001
This compound, (24R)-(+)-5β-cholestane-3α,7α,12α, 24,26,27-hexol, has been designated as 24R-scyrnnol.
The chemical structure of scymnol has been reported by Bridgwater et.al. .Biochem.J. (1962) 8_2 : 285) as 5β-cholestane-3α,7α,12α,24ε,26,27-hexol. However, the stereochemical configuration at the 24-position of scymnol was not identified, and there are three possibilities in the configuration at this position, namely 24R, 24S or a mixture of 24R and 24S. Bridgwater et.al. also reported that chemically synthesized scymnol was exactly identical with scymnol derived from natural shark's bile. This suggests that scymnol is the mixture of 24R and 24S compounds, since theoretically the synthesized scymnol should be produced in both 24R and 24S configurations on reduction with NaBH4 of the ketone in the 24-position of the starting material as described by Bridgwater et.al.
The present invention provides a method for the preparation of the compound of general formula I, which comprises the step of hydrolysis of a 24R-scymnol sulphate ester with an inorganic acid, preferably in the presence of barium chloride. The process for the preparation of 24R-scymnol as broadly outlined above is particularly advantageous as it can be carried out using aqueous acids. In accordance with a preferred procedure of the present invention, 24R-scymnol has been prepared by hydrolysis of sodium scymnol sulphate with dilute hydrochloric acid in the presence of barium chloride, to give a crystalline product. The physical and chemical data of the product are set out in Table 1:
TABLE 1 Physical and chemical data of 24R-scymnol.
mp 183-184°C (190°C)
(colorless plate), H20 insol, MeOH,Etoh sol AcOEt slightly sol, CHCI3 insol
(α)p 25=40.40°(0.5c,ir_MeOH) (34±2°(0.9c,in EtOH) )
High resolution mass; calcd for C27H44θ4(des-2H20) 432.6493 found 432.3247
IR v KBrcrrT:3400, 2950, 1480, 1380, 1080, 1040, 980, 920 max
1H-NMR(in CD3OD) ;S(ppm) : 3.95(lH,br), 3.80-3.50(6H,m) , 3.48(lH,m), 2.40-2.15(2H,m) , 2.10-1.10(23H,m) , 1.07(3H,d,J=6.01Hz)0.91(3H,s), 0.71(3H,s)
13C-N R(in CD3OD) ; S (ppm)7 .8(d) , 73.5(d), 73.0(d), 69.8(d), 62.7(t), 62.0(t), 50.0(t), 49.0(d), 48.1(s), 43.8(d), 43.6(d), 41.7(d), 41.1(t), 37.8(d), 37.2(t), 36.6(S), 36.5(t), 34.0(t), 33.0(t), 31.9(t), 30.3(t), 29.5(t), 28.5(d), 25.0(t), 24.0(g), 18.9(g), 13.8(g). The stereochemical configuration at the 24-position of the compound was determined as R(+) by its crystallographical analysis and the specific optical rotation. The crystal data and atomic parameters of the compound are set out in Table 2 and Figure 1, respectively.
TABLE 2 Crystal data of 24R-scymnol
C27H gOg.CH3OH.H2O, M=518.71, Orthorhombic, Space group P 2-^2, a=18.571(1), b=19.927(2), c=7.984(l)A, V=2954.8A3, Z=4, F(000)=1144, Dc=1.19 g/cm3, Do=1.22 g/cm3, λ(Cu-Kα) =1.54180 A, μ(Cu-Kα)=7.9 cm-1, crystal size 0.2x0.2x0.4 mm.
The activity of 24R-scymnol in the treatment of liver dysfunction has been investigated. In prior
International Patent Application No. PCT/AU87/00281, two assays have been designated to identify characteristic pharmacological activities of the substance, isolutrol. The bioassays, designated as (A) and (B) , have been based on the following activities:
(A) the active principle prevented liver damage in mice caused by carbon tetrachloride; and
(B) the active principle increased the respiration rate in mice when a toxic substance, such as nicotine, was administered.
These assays are useful in ascertaining the existence of activity, however it has been found that they are sometimes unreliable and not reproducible, indicating they are not suitable assays for measuring the degree of activity. 24R-scymnol has some activity in these two bioassays, but it is difficult to make comparison of the degree of activity between isolutrol and 24R-scymnol. Accordingly, a new assay has been designed to reproducibly measure the degree of activity. This bioassay, designated as bioassay (C) , measures tyrosine aminotransferase (TA) activity in liver of mice. TA is one of important enzymes in liver. ASSAY (C): Mice (5 weeks old) were orally administered the active principle of shark's bile (MD) (lOmg/kg), 24R-scymnol (lOmg/kg), or water (6:00 PM on the previous day, 9:00 AM on the day). After one hour from the last administration, the mice were forced to swim in a water at 35°C. After 4 hours swimming, mice were sacrificed by decapitation and livers were perfused with 0.145 M KCl to remove blood. Half gram of liver was homogenized in 0.145 M KCl and centrifuged at 10,000xg for 30min. and TA activity in the supernatant was measured by the method of Diammondstone. The activity is shown as the amount of p-hydroxyphenyl pyruvic acid (p-HPP) produced by the enzyme reaction for lOmin.
The results of the comparison of the activities of the active principle and 24R-scymnol by the assay (C) are as follows:
TA activity (n-mole p-Hpp/mgprotein/lOmin. )
Control 1007+49.6
MD 1322+63.0**
24R-scymnol 1351±29.2***
** : P < 0.01, *** : P < 0.001
The above results indicate that 24R-scymnol has almost the same activity as the active principle of the shark's bile. The present invention also provides a pharmaceutical composition comprising 24R-scyir.no1, together with a pharmaceutically acceptable carrier or diluent therefor. By way of example, 24R-scymnol can be formulated as stable tablets after being mixed as a powder with a known carrier or bulking agent. Such pharmaceutical compositions may be used, for example, for the activation of liver function in the treatment of the diseases of the liver such as hepatitis, nephritis, diabetes, etc. Further details of this invention will be apparent from the following Examples which illustrate the invention without limiting it in any way.
EXAMPLE 1 - Preparation of 24R-scymnol. The active principle of shark's bile (500mg) was dissolved in 7ml of 1% HC1 containing 400mg of BaCl2 and the mixture was heated for 5h at 100°C. After cooling, the resulting solution was extracted three times with 50ml of AcOH-BuOH (1:1). The organic layer was washed twice with H20. Removal of the solvent gave a yellow oil. The resultant residue was dissolved in MeOH and applied to reversed phase HPLC. 50mg of 24R-scymnol was obtained.

Claims

CLAIMS :
24R-scymnol, having the general formula I
Figure imgf000009_0001
2. The compound of claim 1, in substantially pure form.
3. A method for the preparation of the compound of the general formula I as defined in claim 1, which comprises the step of hydrolysis of a 24R-scymnol sulphate ester with an inorganic acid.
4. A method according to claim 3, wherein said hydrolysis is carried out in the presence of barium chloride.
5. A method according to claim 3 or claim 4, wherein said hydrolysis is carried put with hydrochloric acid.
6. A method according to any one of claims 3 to 5 wherein said 24R-scymnol sulphate ester is sodium 24R-scymnol sulphate.
7. A pharmaceutical composition comprising 24R-scymnol, together with a pharmaceutically acceptable carrier or diluent therefor.
PCT/AU1989/000064 1988-02-19 1989-02-16 24r-scymnol, and preparation and use thereof WO1989007607A1 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
DE68925355T DE68925355T2 (en) 1988-02-19 1989-02-16 24R SCYMNOL, PRODUCTION AND USE
EP89902728A EP0401262B1 (en) 1988-02-19 1989-02-16 24r-scymnol, and preparation and use thereof
DK192890A DK167220B1 (en) 1988-02-19 1990-08-14 24R-Scymnol, process for preparing it, and pharmaceutical preparation which comprises it
US08/176,576 US5451575A (en) 1988-02-19 1993-12-30 Method of treating liver dysfunction with 24-R scymnol

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AUPI685088 1988-02-19
AUPI6850 1988-02-19

Publications (1)

Publication Number Publication Date
WO1989007607A1 true WO1989007607A1 (en) 1989-08-24

Family

ID=3772839

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/AU1989/000064 WO1989007607A1 (en) 1988-02-19 1989-02-16 24r-scymnol, and preparation and use thereof

Country Status (14)

Country Link
EP (1) EP0401262B1 (en)
JP (1) JP2716827B2 (en)
AT (1) ATE132500T1 (en)
AU (1) AU609959B2 (en)
CA (1) CA1328104C (en)
DE (1) DE68925355T2 (en)
DK (1) DK167220B1 (en)
ES (1) ES2013025A6 (en)
GR (1) GR1001203B (en)
HK (1) HK88396A (en)
NZ (1) NZ228023A (en)
PT (1) PT89742B (en)
WO (1) WO1989007607A1 (en)
ZA (1) ZA891242B (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992000316A1 (en) * 1990-06-30 1992-01-09 Tsumura & Co., Ltd. Bile alcohol and drug containing the same as active ingredient
CN100383157C (en) * 2006-06-01 2008-04-23 中国科学院南海海洋研究所 3beta, 5 alpha, 6 beta, 11 beta tetrahydroxy cholestanol, and its preparing method and use

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU597723B2 (en) * 1986-08-21 1990-06-07 J.W. Broadbent Nominees Pty. Ltd. Sodium scymnol sulphate isolated form shark tissues
ZA947912B (en) * 1993-10-12 1995-05-24 Broadbent J W Nominees Pty Ltd Treatment of medical disorders associated with free radical formation
IL139241A0 (en) * 1998-05-13 2001-11-25 Novo Nordisk As Meiosis regulating compounds
AU2008275296A1 (en) 2007-07-06 2009-01-15 Sanofi Pasteur Vaxdesign Corporation Rapid generation of T cell-independent antibody responses to T cell-dependent antigens

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3931403A (en) * 1973-05-25 1976-01-06 Intellectual Property Development Corporation Antimicrobial compositions

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3931403A (en) * 1973-05-25 1976-01-06 Intellectual Property Development Corporation Antimicrobial compositions

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992000316A1 (en) * 1990-06-30 1992-01-09 Tsumura & Co., Ltd. Bile alcohol and drug containing the same as active ingredient
CN100383157C (en) * 2006-06-01 2008-04-23 中国科学院南海海洋研究所 3beta, 5 alpha, 6 beta, 11 beta tetrahydroxy cholestanol, and its preparing method and use

Also Published As

Publication number Publication date
ES2013025A6 (en) 1990-04-16
HK88396A (en) 1996-05-31
ZA891242B (en) 1989-10-25
PT89742A (en) 1989-10-04
JPH03503407A (en) 1991-08-01
DK167220B1 (en) 1993-09-20
EP0401262A4 (en) 1993-05-05
AU3186189A (en) 1989-09-06
GR1001203B (en) 1993-06-21
DE68925355D1 (en) 1996-02-15
DK192890D0 (en) 1990-08-14
PT89742B (en) 1994-03-31
CA1328104C (en) 1994-03-29
JP2716827B2 (en) 1998-02-18
EP0401262B1 (en) 1996-01-03
DK192890A (en) 1990-08-14
EP0401262A1 (en) 1990-12-12
ATE132500T1 (en) 1996-01-15
DE68925355T2 (en) 1996-08-01
AU609959B2 (en) 1991-05-09
NZ228023A (en) 1991-10-25

Similar Documents

Publication Publication Date Title
JP2619951B2 (en) Gabapentin monohydrate and method for producing the same
SHIBANUMA et al. Synthesis of optically active 2-(N-benzyl-N-methylamino) ethyl methyl 2, 6-dimethyl-4-(m-nitrophenyl)-1, 4-dihydropyridine-3, 5-dicarboxylate (Nicardipine)
GB2053924A (en) Immunosuppressive 3-hemiesters of7-oxygenated cholesterol
CA1326029C (en) 4-thiazolidine-carboxylic acid derivative, its preparation and pharmaceutical compositions containing it
CA1312090C (en) Hydroxyalkylcysteine derivative and expectorant containing the same
WO1989007607A1 (en) 24r-scymnol, and preparation and use thereof
US5760009A (en) Spirostanyl glycosidal crystalline monohydrate
EP0228625B1 (en) Peptide derivatives inhibitors of hydroxyling enzymes, their process of preparation, the agents thereof and their use
CA2433190C (en) Amlodipine hemimaleate
US5986142A (en) Process for preparing bicycloheptanamine compounds
US5081149A (en) Antihepatopathic method
CA1050996A (en) Antidiabetic agent, &#34;isoxazole derivatives&#34;
CN1128999A (en) Heterocyclic compounds
JPH0678231B2 (en) Blood viscosity reducing agent
WO1980000444A1 (en) Hydroxamic acid derivatives
US4491581A (en) 4-(2,1,3-Benzoxadiazol-4-yl)-1,4-dihydropyridazine derivatives, their production and pharmaceutical compositions
WO1992019612A1 (en) Antiarteriosclerotic agent, a process for the preparation thereof and the use thereof
US3117059A (en) O-benzylphenoxy-
US4034045A (en) 2,4-Disubstituted-4b,5,6,7,8,8a,9,10-octahydro-9-oxo-phenanthrenes
US4051249A (en) Salts of 2-alkylthiazole-5-methanol derivatives
JP2640239B2 (en) Protostar-13 (17) ene-3,16-dione compound
US4097609A (en) 2,4-Disubstituted-4b,5,6,7,8,8a,9,10-octahydro-9-oxo-phenanthrenes
US3598861A (en) 2-(5&#39;-phenyl-m-terphenyl - 4 - yloxy) lower aliphatic monocarbocyclic acids and esters thereof
GB2108966A (en) 25-substituted-5-a-cholestane-3b-22s-diols and esters
JPH05148279A (en) Bisphosphonic acid derivative

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU DK JP KR US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE FR GB IT LU NL SE

WWE Wipo information: entry into national phase

Ref document number: 1989902728

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1989902728

Country of ref document: EP

WWG Wipo information: grant in national office

Ref document number: 1989902728

Country of ref document: EP