JPH03271239A - Triterpene derivative - Google Patents

Abstract

NEW MATERIAL:A compound of formula I (R<1> is H, alkyl, acyl, aralkyl or oligosaccharide residue with 1-3 units; R<2> is methyl, hydroxymethyl or formyl; R<3> is methyl or hydroxymethyl; R<4> is H, alkyl, acyl or aralkyl). USE:A therapeutic or preventive agent for asthma, hypertension etc. PREPARATION:Dry roots of Brazil licorice is extracted with a 80% aqueous methanol solution followed by concentration. Ethyl acetate is then added to the resulting concentrate followed by partition fractionation to recover a water- soluble fraction. Water-saturated n-butanol is then added to this fraction followed by extraction to recover an organic layer, which is then put to fractionation using normal-phase adsorption chromatography, reversed-phase partition chromatography, ion exchange chromatography, or an appropriate combination thereof and others, thereby providing a compound of formula II, etc., which is then hydrolyzed to obtain the objective compound of the formula I where R<1> and/or R<4> is (are) H.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to triterpene derivatives having 3',5'-cyclic nucleotide phosphogesterase inhibitory activity.

[Prior art]

Adenosine-3°, 5′-cyclic monoline # (hereinafter referred to as C-
AMP) and guanosine-3′,5°-cyclic monophosphate are 1-2
It is said that it acts as a ``metsenger'' and is involved in cell membrane function, cell proliferation, and differentiation.
', 5'-cyclic nucleotide is decomposed to form 5
The enzyme that converts the
DE) (EC3,1,4゜17), and substances that inhibit this PDE increase the level of 3',5'-cyclic nucleotides in the body, so they have various physiological effects, such as bronchodilation. It is known to have effects such as cardiotonic action, smooth muscle relaxing action, and hormone secretion promoting action.

As substances that inhibit PDE, xanthine derivatives (methylxanthine, theophylline, caffeine, aminophylline, etc.), papaverine, vinpocetine, milrinone, etc. are known, and some of them are used as cerebral circulation improving drugs.

[The problem that the invention attempts to solve I! ] The present inventors have conducted extensive research from a unique viewpoint apart from the above-mentioned conventional techniques in order to develop a substance that has strong PDE inhibitory activity and is highly specific.

(Means for solving !If) The present inventors have discovered that Periandra duLcis Mart, which belongs to Leguminosae (Fabaceae),
Highly specific and powerful PD from the root of (Brazilian licorice)
Extraction of novel triterpene derivatives with E inhibitory activity,
After completing the separation and identification, various derivatives of the active acid were tested and their PDE inhibitory activity was confirmed.

That is, the present invention provides a method using the general formula (wherein R1 is a hydrogen atom, alkyl, acyl, aralkyl, or an oligosaccharide residue having 1 to 3 sugar units, R1 is methyl, hydroxymethyl, formyl, and R3 is methyl, hydroxy triterpene derivatives (hereinafter referred to as
(referred to as compound (I)).

In the present invention, alkyl preferably has 1 to 6 carbon atoms, particularly 1 to 4 carbon atoms, and may be either linear or branched, and specifically includes methyl, ethyl, n-propyl, isopropyl, and isobutyl. , dobutyl, t-butyl, S-butyl, n-pentyl, isopentyl, neopentyl, t-pentyl, n-hexyl and the like. The acyl may be either aliphatic or aromatic; examples of the aliphatic type include those having 1 to 6 carbon atoms such as formyl, acetyl, propionyl, butyryl, and valeryl; examples of the aromatic type include benzoyl. , cinnamoyl, and those having other substituents on the benzene ring, such as toluoyl, salicyloyl, phthaloyl, syringyl (4-
oxy-3,5-dimethoxybenzoyl) and the like. Furthermore, examples of aralkyl include those in which the above-mentioned alkyl is substituted with aryl such as phenyl, tolyl, xylyl, biphenyl, and the like.

Furthermore, the small amount of 1 residues in sugar units 1 to 3 represented by R3 refers to mono-, di- or tri-glycoside residues. Its constituent line is not particularly limited, and specifically, monoglycoside residues include glucosyl group, arabinosyl group, galactosyl group, mannosyl group, fructosyl group, xylosyl group, ribosyl group, abiosyl group, glucosamine group,
Rhamnosyl group, glucuronosyl group, diglyside residues include abiosylglucosyl group, scrosyl group, maltosyl group, lactosyl group, gentiobiosyl group, xylosylglucuronosyl group, glucosylglucuronosyl group, and triglycoside residues include abiosylgen. Examples include thiobiosyl group, gentianosyl group, rhametsyl group, rhamnosylxylosylglucuronosyl group, rhamnosylglucosylglucuronosyl group, and these structures tcI! It also includes sugar acid and sugar alcohol groups.

This compound (1) can be obtained, for example, as follows.

First, the dried roots of Brazilian licorice are extracted with an 80% aqueous methanol solution, concentrated, and then ethyl acetate is added to perform partitioning and fractionation to collect a water-soluble fraction. This water-soluble fraction has a water-saturated n
- Add butanol, extract and collect the organic layer.

This organic layer was subjected to normal phase adsorption chromatography (silica gel, activated alumina, etc. as a packing material, chloroform + methanol, chloroform + methanol + water, dichloromethane (or chloroform) + methanol + raw acetic acid water, ethyl acetate + ethanol + water, hexane+
acetone, etc.), reversed phase partition chromatography [C3@(octadecylsilane), C, (octylsilane), etc. as packing material, methanol Chiki, acetonitrile plain water, water in tetrahydrofuran, etc. as elution solvent], ion exchange chromatography (QAE-1 as a filler)
Fractionation using an appropriate combination of other chromatographies (Amberlite XAD-2, etc. as a packing material, methanol + water, etc. as an elution solvent); The compound (1-1) represented by the formula or the compound (I-2) represented by the formula is obtained.

These compounds (1-1) or (T-2) are hydrolyzed by a conventional method to obtain a compound (I) in which R1 and/or R4 are hydrogen atoms.

In addition, hydrogen atoms of R1 and/or R4 can be replaced with other substituents (alkyl, acyl, aralkyl, sugar unit 1
~3 minor III residues).

In addition, compound (1-1) or (I-2) is reduced by a conventional method, and R'' is -cuton (hydroxymethyl).
, and further compounds represented by -CHx (methyl) (
I) and a compound (1) in which R3 is -CH3 (methyl) are obtained.

Compound (1) obtained in this way is subjected to dissolution, recrystallization,
Isolated by conventionally known methods such as chromatography,
Can be purified.

The drug containing the compound (1) of the present invention as an active ingredient can be used as a treatment/improvement drug for specific diseases in mammals (humans, horses, dogs, mice, rats, etc.), such as a therapeutic and preventive agent for asthma, hypertension, etc. It is extremely useful clinically.

The drug containing the triterpene derivative of the present invention as an active ingredient is
It can be administered orally or parenterally, but in the case of oral administration, compound (1) is mixed with appropriate pharmaceutically acceptable additives (carriers, excipients, etc.).
diluents, etc.) and used as scatterings, tablets, capsules, troches, solutions, syrups, and granules. In the case of parenteral administration, it is used as an aqueous solution or non-aqueous suspension, an injection such as intravenous injection, intramuscular injection, subcutaneous injection, cross preparation, etc.

The dosage may vary depending on the patient's symptoms, weight, age, etc.

〔effect〕

The compound CI) of the present invention has high specificity (particularly against PDE-■) and has strong PDE inhibitory activity, so it can be used clinically as a treatment/improvement drug for specific diseases, such as asthma and hypertension. 6 [Example] (Example 1) 8 R' = -0-heavy
Luron 2-1 Xylose t ~ 1-mu -se R2 Extract 146.6 kg of dried Brazilian licorice with 80% methanol aqueous solution, concentrate, add ethyl acetate to partition, and collect the water-soluble fraction. . Water-saturated n-butanol is added to this water-soluble fraction for extraction and the organic layer is collected.

This organic layer was transferred to the C1 column at 35% as a mobile phase solvent.
Fractionation was performed by preparative reverse-phase high performance liquid chromatography (HPLC) using an acetonitrile aqueous solution. On this occasion,
The flow rate was set to 20 (ld/■in), and fractions were taken every 5 minutes, and the fractions between 20 and 30 minutes were collected.

Next, dichloromethane-methanol-
Acetic acid-water (60-80: to 30:5-20:1)
5) Preparative normal phase HPLC or silica gel column chromatography using mixed solvent and C, column or 0DP
(Product name, manufactured by Asahi Kayi Co., Ltd., polymer-based 04 column) Fractionation by both preparative reverse-phase HPLC using 35% acetonitrile aqueous solution as the mobile phase solvent in the column was performed to determine PDE inhibitory activity. As an indicator, compounds (1-1
) 14.5■ was obtained.

The physical properties of this compound (1-1) were as follows: Optical rotation Ccx] m''-55,0 (C=0.2, methanol) 'H-NMR (500,135 MHz, pyridine-d2
．． Temperature: 297 K) 0.90 H (Example 2) In Example 1, a part of the fluxian after compound (1-1) was eluted by the first preparative reverse-phase HPLC was used as a preparative sample. Fractionation and purification were performed by reverse phase HPLC (35% acetonitrile aqueous solution), and compound (I-2) was
■Isolated.

The physical properties of this compound (1-2) were as follows: Optical rotation [α] 1°--17.4°C (C = 0.5, pyridine) 'H-NMR (200,133MHz, pyridine- d3
．． The mixture was dissolved in sulfoxide (temperature: 323 K), and 200 μ of water (5-1) sodium borohydride was added thereto, followed by stirring at room temperature for 62.5 hours.

After post-treatment using a conventional method, preparative reverse-phase HPLC (35
% acetonitrile aqueous solution) to obtain the compound (1-
3) Obtained 6.3■.

Optical rotation [α] D”=29.0 (C=0.5.
pyridine) 'H-NMR (200,133MHz, pyridine-d
3. Temperature: 297 K) (Example 3) (Example 4) 20 g of compound (1-2) was dissolved in DMSO (dimethyl compound (
1-2) To the solution of 20μ in diethylene glycol 2d, add potassium hydroxide (700μ) in diethylene glycol solution (20m) and 80% hydrazine hydrate 2
0d was added thereto, and the mixture was heated under reflux on an oil bath (150 to 160°C) for 54 hours.

After distilling off excess hydrazine and water, the mixture was heated under reflux (oil bath temperature: 200° C.) again for 12 hours.

After cooling, 10% sulfuric acid was added to adjust the pH to approximately 2, extraction was performed with ethyl acetate, and the ethyl acetate layer was washed and washed in a conventional manner.
After drying, Na was reduced to dryness under reduced pressure.

The residue was purified by preparative normal phase HPLC (isopropyl alcohol:hexane = 119) to obtain compound (I-
4) Obtained 1.0■.

Heavy H-NMR (200,132 MHz, heavy chloroform, temperature 297 K) The PDEvA harmful activity of the active substance was determined by specific activity.

Using three types of PDEs derived from bovine heart that were fractionated by DEAE-cellulose column chromatography, PD
The 50% inhibitory concentration (μM) of each compound against H was measured. Note that 1 μM of c-AMP was used as a substrate.

The results are shown in Table 1 below.

In Table 1, the value of milrinone is a literature value, and the three types of PDE used were those derived from guinea pig heart.

Regarding compound (1-1), furthermore, Dixson (Dix
On) When we investigated the competitive inhibition mode by plotting, we found that K
! =0.063 μM.

[Test example]

Claims (1)

[Claims] (1) General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (I) (In the formula, R^1 is a hydrogen atom, alkyl, acyl, aralkyl, oligosaccharide residue of sugar units 1 to 3, R^2 is methyl,
A triterpene derivative represented by hydroxymethyl or formyl, R^3 represents methyl or hydroxymethyl, and R^4 represents a hydrogen atom, alkyl, acyl, or aralkyl.