JPH0393727A - Blood flow improver - Google Patents

Abstract

PURPOSE:To obtain a blood flow improver having durable and strong effects free from side effect, comprising Epidermal Growth Factor(EGF) or a salt thereof as an active ingredient. CONSTITUTION:A blood flow improver containing EGF such as beta-urogastrone (hEGF1-53) having an amino acid sequence shown by the formula (dotted line is disulfide bond), beta-urogastrone (hEGF1-52) which is deficient in one amino acid from C end of gamma-urogastrone or other EGFs wherein these constituent amino acids are partially replaced with other amino acids or pharmaceutically acceptable salt thereof. The EGF is produced by extraction from human urine and purification or gene recombination technology.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention provides epidermal growth factor (Epiderm-al
The present invention relates to a blood flow improving agent containing Growth Factor (hereinafter abbreviated as EGF) or a pharmaceutically acceptable salt thereof as an effective component.

[Conventional technology]

In 1975, H. Gregory purified a peptide factor that strongly inhibits gastric acid secretion from human urine (named it β-urogastrone) and determined the entire 53 amino acid sequence (Nature, Vol. 257). ,3
25-327, 1975).

On the other hand, apart from Gregory's research, S. Cohen et al. investigated a peptide that promotes eyelid cleavage in newborn mice, which is abundant in mouse submandibular glands, from the perspective of a factor that promotes the proliferation of epithelial cells. He named it EGF, studied its physiological effects and structure, and in 1972 elucidated the entire amino acid sequence (53 amino acids) of mouse EGF (J, Bi
o l, C. hem, vol. 247, 7612-76
21, 1972).

Furthermore, they discovered that human EGF has the same physiological effects as mouse EGF.
We discovered that EGF (hereinafter abbreviated as hEGF) is also present in the urine of pregnant women, and showed its amino acid composition (Pro
c. Nat. Acad. Sci. USA, vol. 72, 13
17-1321, 1975).

Today, hEGF and β-urogastrone are the same substance.
It is widely accepted that the amino acid sequence of the following formula (I) determined by Gregory is correct for its structure.

The physiological effects of EGF (in the blanks below (- indicates a disulfide bond) include the above-mentioned effects of suppressing gastric acid secretion and promoting cell proliferation, as well as promoting corneal repair (Savage, C).
．． , et al.: Exp t. Eye Res. ,15
Vol. 361, 1973; JP-A-59-65020), bone repair promoting effect (Kumegawa, H.,
et al., Calcif. Tissue Int. 35
Volume, 542 pages, 1983; JP-A-61-293925
Publication No.) etc. are known.

By the way, the organs and tissues of a living body maintain homeostasis by being supplied with oxygen and nutrients through the bloodstream and at the same time removing carbon dioxide gas, metabolic products, etc. through the bloodstream. Therefore, for the living body to function properly, an appropriate amount of blood flow must be maintained at all times. Blood flow is impaired by various causes, such as cardiac dysfunction, disorders of the vascular system (vasoconstriction, arteriosclerosis, thrombosis, etc.), autonomic nerve dysfunction, and the like. Conventionally, blood flow improving agents include those that act systemically such as vasodilators such as propranolol and verabamil, and those that act locally such as gastric mucosal blood flow improving agents such as prostaglandin El and sofalcon. Cerebral blood flow improving agents such as calcium hopatenate and citicoline are known.

[Problem to be solved by the invention]

However, the problem with these blood flow improving agents is that they have strong side effects when administered systemically and require great care when administered, and when used locally they have a weak blood flow improving effect and their effectiveness is uncertain. be. An object of the present invention is to provide a blood flow improving agent that has few side effects and is highly effective.

[Means to solve the problem]

In order to achieve the above object, as a result of various studies, it was unexpectedly discovered that EGF has a sustained and strong blood flow improving effect, and the present invention was completed.

That is, the present invention relates to a blood flow improving agent containing epidermal growth factor (EGF) or a pharmaceutically acceptable salt thereof as an active ingredient.

EGF used in the present invention may have the same physiological activity as β-urogastrone (hEGFx-s3) shown in formula (1), although there may be differences in degree. As such, γ-urogastrone (h E
G Fl-52) and hEGFt-n with 2 to 7 deletions
(n is an integer from 51 to 46), some of these constituent amino acids are substituted with other natural or unnatural amino acids, some of the constituent amino acids are deleted or added, etc., and mixtures of these But that's fine.

These EGFs can be produced by various methods, such as the method of extraction and purification from human urine (Gregory et al.
, H., et al., Hoppe Sey-Iers.
Z. Physiol, Chem, vol. 356, 17
65-1774, 1975; Gregory, H.
, Nature, vol. 257, pp. 325-327, 197
5 years; Japanese Patent Application Publication No. 58-99418; 58-219
No. 124; No. 60-161923, etc.), a method using genetic engineering technology (Japanese Unexamined Patent Publication No. 57-122096)
No. 58-216697; No. 59-132
No. 892, etc.).

The blood flow improving agent of the present invention includes a formulation consisting of a pharmaceutically acceptable carrier or diluent and EGF or a pharmaceutically acceptable salt thereof. Preferred examples of the salts include metal salts such as alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts and magnesium salts, ammonium salts, organic bases, organic acid salts, and inorganic acid salts. Can be mentioned.

The formulations can also be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to a patient.

Forms of administration include injections, oral drugs, inhalers,
Take suppositories etc. as appropriate. Typical methods of administration include oral, rectal, percutaneous, subcutaneous, intravenous, intramuscular, inhalation, or intranasal routes.

In these administration methods, the blood flow improving agent of the present invention can be administered in the form of various pharmaceutical preparations. The forms of these pharmaceutical preparations include tablets, hard capsules, soft capsules, granules, powders, troches, suppositories, syrups, creams, ointments, poultices, injections, suspensions, There are inhalers, aerosols, etc. Also, double layer tablets, along with other medicines.
It can also be a multi-layer tablet. Furthermore, in the case of tablets, they can be coated with a conventional coating, for example, sugar-coated tablets or enteric-coated tablets.

When preparing solid preparations such as tablets, granules, and powders, known additives such as lactose, sucrose, glucose, crystalline cellulose, corn starch, calcium phosphate, sorbitol, glycine, carboxymethylcellulose, and hydroxybrovir are added to the formulation. cellulose, gum arabic,
Polyvinylpyrrolidone, polyethylene glycol, magnesium stearate, talc, etc. can be added.

In the case of semi-solid formulations, materials such as vegetable waxes, microcrystalline waxes, fats such as tarolanoline, etc. can be added.

When preparing a liquid formulation, additives such as sodium chloride, sorbitol, glycerin, olive oil, almond oil, brobylene glycol, ethylene glycol, and ethyl alcohol can be added.

The dose of EGF can be increased or decreased as appropriate depending on the patient's age, weight, symptoms, etc., but it is usually 0.01 per dose.
~100 mg/kg.

〔Example〕

The present invention will be explained in more detail with reference to Examples described below. The test drug, EGF, was isolated and purified from healthy human male urine, and was derived from β-urogastrone (hEGFx).
-s3) 14%, γ-urogastrone (h E G F
l-52) 6 7%, in addition hEGFl-49, h
Those containing small amounts of E G Ft-so and h E G Ft-sl were used.

(Example 1) A male Wista strain rat weighing approximately 300 g was subjected to laparotomy under anesthesia, a small hole was made in the forestomach, a laser probe was inserted into the stomach, and gastric mucosal blood flow was measured using a laser Doppler blood flow meter. Measured over time. As shown in Figure 1, the test drug EGF (dissolved in physiological saline) was intravenously administered at 20 μg/140 μg/1 kg of rat body weight, and 0.5 ml of physiological saline as a control was administered intravenously at the point indicated by the arrow. Mucosal blood flow was measured for 90 minutes. The results shown in FIG. 2 are the average of five repeated experiments, and the blood flow is expressed as a relative value, with the blood flow at the time of administration of the test drug being taken as 100.

From the results in Figure 1, EGF is 20μg/kg and 40μg/kg.
It was found that administration of the drug at a dose of 1.5 g/kg had a sustained effect of increasing blood flow for about 90 minutes after administration.

(Example 2) An incision was made on the left flank of an adult male mongrel dog weighing 10 to 15 kg under anesthesia, the left renal artery was exposed, a probe was attached, and the left renal artery blood flow was continuously measured using an electromagnetic blood flow meter. It was measured.

As shown in Figure 2, the test drug EGF (dissolved in physiological saline) was added to the dog at the point indicated by the arrow.
5 μg/kg, 110 μg, 20 μg, and 0.5 ml of physiological saline as a control were intravenously administered, and the blood flow was measured for 30 minutes. The results shown in FIG. 2 are the average of four repeated experiments.

From the results shown in FIG. 2, EGF can significantly increase renal blood flow for about 20 minutes when administered at a dose of 5 μg/kg or more.

〔Effect of the invention〕

According to the present invention, it is possible to provide a blood flow improving agent that has few side effects and is highly effective.

[Brief explanation of drawings]

Figure 1 is a graph showing the effect of EGF on gastric mucosal blood flow in rats, and Figure 2 is a graph showing the effect of EGF on renal artery blood flow in dogs.
It is a graph showing the effect of F.

Claims (1)

[Claims] 1. A blood flow improving agent containing epidermal growth factor or a pharmaceutically acceptable salt thereof as an active ingredient.