JPH05117142A - Lipid depressant containing hopanoid - Google Patents

Abstract

PURPOSE:To obtain a lipid depressant with excellent lipid-depressant effect by using a substance containing hopanoid which is represented by a specific formula and has the action to inhibit the secretion of extremely low-density lipoprotein from hepatocytes. CONSTITUTION:Hopanoid (a pentacyclic triterpenoid with a sterol-like structure having a basic skeleton of formula I and distributing in cells and some kinds of plants), for example, bacteriohopan-32-ol is used as an active ingredient to give the objective lipid depressant. The depressant contains the hopanoid, in the form of a preparation suitable for oral or parenteral administration, 1.0 to 100wt.%, preferably 1.0 to 60wt.% and is used for hypercholesterolemia, hyperlipemia, arteriosclerosis. By the way, Hopanoid can strongly inhibit the secretion of apolipoprotein B which is the main protein component of extremely low-density protein and phospholipid with no influence on lipid synthesis and inhibits triglyceride secretion, too.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to the field of medicine.
More specifically, it relates to a lipid lowering agent characterized by containing a hopanoid.

[0002]

2. Description of the Related Art In recent years, ischemic heart disease has become the second leading cause of death after cancer. The main cause of ischemic heart disease is hyperlipidemia. In recent years, the number of patients with hyperlipidemia tends to increase due to changes in diet. As a therapeutic drug for hyperlipidemia, since HMG-CoA reductase inhibitor shows a strong cholesterol lowering effect at present,
It has become popular.

An HMG-CoA reductase inhibitor inhibits mevalonate synthesis by HMG-CoA reductase in the liver, reduces cholesterol in cells, increases the number of LDL receptors on the liver surface, and reduces blood cholesterol. Let However, cholesterol is an essential substance for the living body such as regulation of the fluidity of biological membranes, precursors of steroid hormones, raw materials of bile acids, and HMG-Co.
Mevalonate, which is a reaction product of A reductase, is used as an important component for the living body such as dolichol, ubiquinone, and farnesyl pyrophosphate. Therefore, a drug that exhibits an anti-hypercholesterol action by another action is preferable to a drug that inhibits the cholesterol synthesis system.

[0004]

[Problems to be Solved by the Invention] Creation of a novel blood lipid lowering agent that overcomes the drawbacks of HMG-CoA reductase.

[0005]

Means for Solving the Problems Blood lipids are released from the liver in the form of very low density lipoprotein and are used in various parts of the body. Therefore, the present inventors considered that a drug that inhibits the secretion of ultra-low density lipoprotein from hepatocytes would be an ideal therapeutic drug for hyperlipidemia, and conducted a search for such a drug, and found that the amount of hopanoid was extremely low. The present invention has been completed by finding that they strongly inhibit the secretion of density lipoprotein.

[0006] The hopanoid has the formula

[0007]

[Chemical 1] It is a pentacyclic triterpenoid with a sterol-like structure that exists in bacteria and certain plants, and has a basic skeleton represented by, and its related compounds are also widely present in sediments on the earth [Annual Review of Microbiology (Ann. Rev. Microbio
l. ), 41, 301, 1987]. In addition, various related compounds have been synthesized by chemical modification [J. General Microbiology,
131, 1363, 1985].

[0008] It is considered that hopanoids present in prokaryotes such as bacteria play a role similar to that of sterols in the biomembrane of eukaryotes. On the other hand, cholesterol is involved in the regulation of cholesterol biosynthesis in the living body, and when the cholesterol in the living body increases, HMG-CoA reductase, which is the rate-limiting enzyme of the cholesterol biosynthesis system, is feedback-inhibited and the amount of cholesterol synthesis decreases.
It is believed that oxysterols, not cholesterol, are directly involved in this down regulation. Especially 25-
Hydroxycholesterol in cells is HMG-CoA
It is known to down-regulate reductase and reduce cholesterol synthesis. Hopanoids are similar in structure to cholesterol and oxysterols. Therefore, we examined the effect of hopanoid on lipid synthesis and ultra-low density lipoprotein secretion, and investigated the secretion of apolipoprotein B and phospholipid, which are the major protein components of ultra-low density lipoprotein, without affecting the lipid synthesis. It was found that it strongly inhibits and also inhibits triglyceride secretion. Hopanoids are the first substances other than hormones to suppress the secretion of apolipoprotein B under these conditions.
The hopanoid found in the present invention is a known substance, but the present inventors were the first to find that the hopanoid strongly suppresses apolipoprotein B secretion and phospholipid secretion and also inhibits triglyceride secretion. .

Hopanoids are useful as lipid lowering agents
This will be shown by the following pharmacological tests. Pharmacological test (1) Action of hopanoid on lipid synthesis and secretion (A) Test compound: Bacteriophane having the following structure
-32-All [Journal of General My
Crobiology (J. General Microbi
method), 131, 1363, 1985.
Manufactured in. ]

[0010]

[Chemical 2] (B) Test method The method used by the present inventors to study lipid synthesis and secretion is as follows.

Lipid synthesis from [ ¹⁴ C] acetic acid or [ ³ H] glycerin is performed by the method of Brown et al. [J. Biol.
m. ), 253, 1121, 1978]. On day 0, 1.2 × 10 ⁵ Hep G2 cells were added to 1
Medium A containing 0% fetal bovine serum (100 units / ml penicillin and 100 μg / m in Eagle's modified minimal essential medium)
(medium to which 1 streptomycin was added) was added.
Sow into a plastic petri dish of cm ² . Replace medium on day 3 or 4 with fresh one. On day 6, replace with medium A without fetal calf serum. After changing the medium on the 7th day, the cells were mixed with Bacteophopan-32-ol for 14 days.
Pre-incubate for hours and label with 1 mM [ ¹⁴ C] acetic acid or 2.5 mM [ ³ H] glycerin for 4 hours. After the incubation, cool the plastic dish with ice and collect the medium. Cells are washed 3 times with cold phosphate buffered saline. Cell homogenates or lipids in the medium are analyzed by the method of Forti et al. [J. Biol.
m. ), 226, 497, 1957].
When [ ³ H] glycerin was used as a precursor, phospholipids and triglycerides were separated on a silica gel G plate using hexane / diethyl ether / acetic acid (85: 15: 4, V / V) as a developing solvent. When [ ¹⁴ C] acetic acid is used as a precursor, phospholipids, free cholesterol,
Triglycerides and esterified cholesterol are hexane / diethyl ether / methanol / acetic acid (85: 1
5: 1: 1, V / V) as a developing solvent. Thin layer chromatography (TLC) plates are developed twice to completely separate lipids. The lipids on the TLC are confirmed by exposure to iodine vapor. The radioactivity in the area corresponding to the standard pure product is measured with a liquid scintillation counter. The protein measurement was carried out by the method of Lowry using bovine serum albumin as a standard substance [Journal of Biological Chemistry (J. Biol. Chem.), 1].
94, 265, 1951].

As shown in Tables 1 and 2 below, bacteriophane-32-ol strongly inhibited the secretion of phospholipids and the secretion of triglycerides without affecting the lipid synthesis.

[0013]

[Table 1]

[0014]

[Table 2] (2) Effect of hopanoid on secretion of apolipoprotein (A) Test compound: bacteriophopan-32-ol (B) Test method On day 0, 1.2 × 10 ⁵ Hep G2 cells were added to 3.8.
cm ² containing medium 10% fetal bovine serum (Eagle modified minimal essential medium 100 units / ml of penicillin and 10
Plate on a plastic Petri dish containing 0 μg / ml streptomycin). Replace medium on day 3 or 4 with fresh one. On day 6, replace with medium A without fetal calf serum. After incubating the cells with Bacteophopan-32-ol on the 7th day for 18 hours, the plastic dish is cooled with ice and the medium is collected. Apolipoprotein B secreted into the medium was analyzed by enzyme immunoassay [Clinical Chemistry (Clinica).
Chemistry), 32, 1484, 198.
6].

As a result, 20 and 40 μm bacteriophopan-32-ol inhibited the secretion of apolipoprotein B by 15% and 20%, respectively.

As described above, hopanoid has an excellent lipid-lowering action.

The drug of the present invention, which is characterized by containing a hopanoid, can be orally or parenterally administered, and by formulating into a form suitable for such administration, hypercholesterolemia, It can be used for treatment and improvement of hyperlipidemia and arteriosclerosis. In clinical use of the compound of the present invention, it is also possible to add pharmaceutically acceptable additives in accordance with the dosage form and administer it after various preparations. As an additive at that time,
Various additives usually used in the pharmaceutical field can be used, for example, gelatin, lactose, sucrose, titanium oxide,
Starch, crystalline cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose, corn starch, microcrystalline wax, white petrolatum, magnesium metasilicate aluminate, anhydrous calcium phosphate, citric acid, trisodium citrate, hydroxypropyl cellulose, sorbitol, sorbitan fatty acid ester, polyvinyl Examples thereof include pyrrolidone, magnesium stearate, light anhydrous silicic acid, talc, vegetable oil, benzyl alcohol, gum arabic, propylene glycol or polyalkylene glycol.

Dosage forms formulated as a mixture with these additives include solid preparations such as tablets, capsules, granules, powders or suppositories, or syrups, for example.
Liquid preparations such as elixirs and injections are available, and these preparations can be prepared according to the usual methods in the field of preparation. The liquid preparation may be dissolved or suspended in water or another suitable medium at the same time. Further, particularly in the case of an injection, it may be dissolved in physiological saline or glucose solution, if necessary, and a buffer or a preservative may be added.

These preparations contain the compound of the present invention in an amount of 1.0 to 100% by weight, preferably 1.0 to 60% by weight, of the total drug.
It can be contained in the ratio of. These formulations may also contain other therapeutically effective compounds.

When the compound of the present invention is used as an antihyperlipidemic agent, an antiarteriosclerotic agent or an antihypercholesterolemic agent, the dose and frequency of administration are the sex of the patient, age, weight, degree of symptoms and degree of symptoms. Generally, in the case of oral administration, 0.01 to 20 mg / kg is divided into 1 to several times per day for an adult, and in the case of parenteral administration, it is 0 depending on the type and range of the desired therapeutic effect. 0.001 to 2 mg / kg is 1
It is preferable to administer the drug in several divided doses.

[0021]

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples. Example 1 25 parts of Bacteophopan-32-ol, 70 parts of lactose,
After uniformly mixing 30 parts of corn starch, 23 parts of crystalline cellulose and 2 parts of magnesium stearate,
Tablets are obtained by a conventional method to give tablets each containing 25 mg of the active ingredient. Example 2 25 parts of Bacteophopan-32-ol, lactose 125
Parts, 45 parts of corn starch and 5 parts of magnesium stearate are uniformly mixed, and then 200 mg is filled into a hard gelatin capsule (No. 2), and the active ingredient 2 is contained in 1 capsule.
A capsule containing 5 mg is obtained. Example 3 50 parts of bacteophopan-32-ol, 700 parts of lactose and 230 parts of corn starch were uniformly mixed, and then 20 parts of hydroxypropyl cellulose and a paste prepared from purified water were added and kneaded to prepare a conventional method. Grain 1g
Inside, granules containing 50 mg of the active ingredient are obtained.

[0022]

EFFECTS OF THE INVENTION The pharmaceutical preparation of the present invention, which is characterized by containing the hopanoid, is expected to be used as a lipid lowering agent.

[0023]

Claims (1)

[Claims] 1. A lipid-lowering agent, which comprises a hopanoid.