JPS6016934A - Antineoplastic agent - Google Patents

Abstract

PURPOSE:To provide an antineoplastic agent containing L-carnosine or its salt as an active component, having high safety, and giving little burden to the liver. CONSTITUTION:The objective antineoplastic agent contains L-carnosine of formula (melting point, 250 deg.C; [a]<20>D=+20.0 deg. (H2O); tasteless and odorless; soluble easily in water; white crystalline powder) as an active component. An extremely excellent effect can be expected in the remedy of gastric cancer, rectal cancer, mammary cancer, uterine cancer, oral cancer, esophageal cancer, cystic cancer, etc. by single use, or in the promotion of the effect of immunopromoting agent such as OK-432 by the combined use with the agent. Dose: 0.5-3g daily by oral administration. It can be administered orally or parenterally in the form of injection, powder, granule, tablet, capsule, enteric tablet, ointment, suppository, etc. L-Carnosine is a dipeptide composed of L-histidine and beta-alanine, and is present abundantly in the skeletal muscle of memmals.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an anti-ulcer agent containing L-lunosine otakeso salt as an active ingredient.

The development of tumor therapeutic agents is based on two broad concepts. One of them is based on the idea that cancer can be suppressed by inhibiting the active nucleic acid biosynthesis of tumor silk. In Japan, for example, pleomycin (Nippon Kayaku Co., Ltd.), mitomycin c I: MMC (Wi + Wafunko Co., Ltd.)], 5
-FU (Kyowa Hakko Co., Ltd.) is a therapeutic agent created based on the ideas of Togato. The second is a non-specific active immunotherapy that uses the host's immunity, based on the idea of treatment with so-called immune stimulants, and is based on the idea of treating with a so-called immune stimulant. Vaccine (SSM (Zeria Pharmaceutical Co., Ltd.)), Crestin (ps
K (Kureha Chemical Industry Co., Ltd., Sankyo Seisakushu Co., Ltd.)] and other therapeutic agents have been widely praised.

However, therapy based on erosion is also completely clinical purpose'f.
There is a flaw that cannot be delayed. In the former case, since the inhibitory effect on nucleic acid synthesis is not specific to cancer, it also inhibits nucleic acid synthesis in areas other than tumor tissue, making it difficult to avoid side effects.

The latter therapy with immunostimulants does not produce an immune response that is cancer-specific, and regardless of the nature and quality of the immunostimulant used, the basic problem is that the immune response produced is not specific to the reticuloendothelial system. is limited by the response function of In order to suppress the rapidly proliferating 1j11 tumor, some means must be devised to increase the efficiency of the immune response. One idea to compensate for this deficiency in immunotherapy is to actively immunize human cancers from other animals and extract immuno-RNA'f from animal tissue.
This is a method to perform specific passive immunization by transferring I to the patient (1α).This method allows the immune RNA produced in other animals to be stored and then administered in large quantities to KC. Although it is possible to obtain an effect that exceeds the entire immune response function unique to the organ, there are many remaining problems such as viral infection, and clinically it is a current problem.
It is conceivable to take measures such as opening the threshold of the tumor-specific immune response function of the reticulum viscera, or opening the entire efficiency of the immunoresponsive cells. Histidine is a dipezotide consisting of L-histidine and β-alanine, discovered in Liebig's meat extract by Gulewltsch in 7900, and is contained in large amounts in the skeletal muscles of mammalian animals. .

Since its discovery, many researchers have conducted research into its physiological significance and pharmacological usefulness.
It remained unresolved to this day.

The present inventors believe that fosocarnosine, or L-histidinyl-γ-aminobutyric acid, was used in the experiment aDDY-3arcoma/
Ni0 (Komiyama Preserved Stock, Kitasato Research Institute Cancer Department) and BA
LB/C-METH-AC Kitadai strain, stored at Daiichi Pharmaceutical Co., Ltd. Central Research Laboratory) was found to be effective against the system.

These two cancer strains are highly proliferative JJC cancer strains that cannot be easily suppressed by immunostimulants (i-). The fact that it effectively acts on both systems proves that there is a possibility that it can be extremely effectively suppressed. We filed an application for a bamboo license as a patent agent (Patent Application No. 5g-1983).

As a result of further intensive research, inventor C) found that monocarnosine, which has one less carbon in the side chain than homocarnosine, has a similar antitumor effect, and completed the present invention.

L-carnosine has %i 1 point, 2SO℃ (decomposition), [α]
Otsu0=+20.0° (H2O), and is a white crystalline powder that is tasteless and odorless and easily dissolves in water. It is represented by the following chemical structural formula, and the pH of the aqueous solution is g, θ ~ g, S.

L-carnosine is a substance (approximately 0.1-0.3%) that is present mainly in the skeletal muscles of various mammalian animals, and is ingested as food from daily meat, and is an essential amino acid - a source of histidine. It is. It is also biosynthesized from L-histidine and β-alanine. After absorption, L-carnosine that is not ingested is broken down into L-histidine and β-alanine by carnosinase and becomes nutrients, and some of it is resynthesized into L-carnosine [an intermediate in carnosine biosynthesis].
β-alanyl-/-methyl-histidine (Anser
There is a lns).

As mentioned above, carnosine has a safety similar to that of food, but has a t1gJ negative and p1 absorption is broken down by carnosinase present in various organs, so many other medicines have a hepatic Il. He was expelled and his liver function deteriorated.
It is a different kind of Confucianism.

Next, the acute antecedents of L-carnosine will be described.

Various doses of carnosine were administered intraperitoneally and orally to 10 acutely toxic mice 'e/group, and symptoms of acute toxicity were observed by FA at S hours after the first administration. LD5o is calculated based on the number of deaths after 72 hours.
erden) method. L-carnosine was prepared by adjusting the volume of administration solution to Q, 7 to 0.3 ml//θ1.
Dissolved in saline.

The symptoms of L-carnosine are 15. θθ0m evening/
Intraperitoneal administration of kP (LD,...) 8 From around 30 minutes, the patient's spontaneous movements decreased and the patient took a prone position, and the respiratory rate decreased and became irregular, but no loss of righting reflex or withdrawal reflex was observed. Occasionally, half of the patients experienced symptoms of twitching and clonic convulsions. As the symptoms progress further, the rollover will occur again, and convulsions will be induced in response to contact stimulation, leading to tonic convulsions t1. The disease progressed to death. 1
Half of the cases died after 30 minutes, rc and go% after ko, and all cases died after eζ. / S, 000my71q
y) Almost no sac was observed after oral administration, but 72
After hours, 1 of the IO cases died.

Table 1 Acute toxicity (72 hours) to DO male mice is as shown in the table, and it can be said that L-carnosine Ild is a compound with extremely low toxicity.

-f7? -L-Carnosine is a substance whose safety has already been established, as it has been developed for about 10 years and has been commercialized as a drug for treating anorexia by Risa in Spain. Also, the ineffective dose of L-carnosine against transplanted cancer is 1 per mouse, that is! ;Om9/kVT:Se) However, the dough has acute toxicity marks due to intraperitoneal administration as described above,
. ? , Og7■/kPv)/ //g/, so the safety of Ichiriki Lunosine is fully measured as ■(.

The method for synthesizing L-carnosine is well known.
alof Bloglcal Chamlstry
,, / Q g, 7! ;3,193g), converting carbobenzoxyβ-alanine to chloride with phosphorus pentachloride,
L-carnosine can be obtained by leading to a methyl ester with methanol, supplying hydroazide to form an azide, coupling it with L-histidine methyl ester, and removing the benzoxy group of cal by catalytic reduction. The present invention also encompasses a therapeutic agent comprising a salt of L-carnosine, including a salt of L-carnosine and 1. There are acid addition salts between carmine moiety and group salts and pharmacologically acceptable acids based on amino groups.
There are salts based on both acid and amino groups. Kal day?
Salts based on acid groups include salts with metals such as sodium, potassium, calcium, magnesium, zinc and aluminum, ammonium salts and substituted ammonium salts, salts with trialkylamines and other amines such as triethylamine; Salts based on amino groups include hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, gluvionic acid, lactic acid, tartaric acid, citric acid, succinic acid, maleic acid, benzenesulfonic acid, toluenesulfonic acid, inorganic acids, and organic acids. However, the free salt of carnosine was chemically calculated using a method known to those skilled in the art.
can be produced by reacting with a selected acid or base.

Next, I will give an experimental example and show that L-carnosine is superior to anti-Pr4.
Explain the tumor effect.

Experimental example materials: DDY mouse, male S week old (obtained from Shizuoka Prefecture Laboratory Animal Agriculture Cooperative Association) Tumor cells: Sarco? / g O (Sarcoma /
ZaO) (Komiyama Preserved Strain, Kitasato Institute FLH Classroom) Transplant:
s X/o' were implanted subcutaneously in the midscapular region
111+ (1 hour after cells were separated from the tumor) Administration: For the control one, saline n, x water 0, /
me'frAIr! =fM It/j doses were administered 7 times every other day from lLtg time (2 days) after transplantation.

In Experiment W, L-carnosine alone was 0. /1
1 carnosine/mσ total in nl physiological saline, 1 carnosine/mσ total in saline, 1 carnosine in 1 me raw milk solution 0 ”-2, /% 2”; Approximately 2 ON 1 liters were administered subcutaneously on the back from the transplanted site, 7 times every other day from 1 hour (2 days) after the tumor was transplanted, for a total of 1. I threw 1 S box. 0 to 1 da? As for Yu, both the Japanese and the combined use are θ, /ml physiological * OK in salt water - 0.3KE of 32
intraperitoneally, 1g and 11f after l11-'I tumor transfer 4j'
The drug was administered 3 times every other day starting from day 1.

(1) Average number of days alive #9 is (each number of days alive x lu of that number of days)
It is expressed as the sum of the raw comparisons) divided by the total comparison.

Since the cases of extinction [7] were not added to the average survival days, young buds are considered in relation to the average survival days and these cases.

(3) "Disappearance" means that the M11 cells in the transplanted lung were separated and clearly grew macroscopically with Mi<1M IIC, but they began to regress midway, and the calluses fell off, making it difficult to distinguish them from control animals. <I say many things. The cases in which the tissue disappeared in the table were indistinguishable from normal tissue at autopsy and histological examination 60 days later.

* Two of these cases are non-engrafted cases (non-engraftment refers to cases where the graft does not grow properly after transplantation and microscopically disappears from the transplanted site).

(4) The size of the tumor was determined by the major axis x the minor axis (bait 2).

In Figures 1 and 2 of the salary, the horizontal axis shows the entire experimental period (day 1 to day 60), and the vertical axis shows the surviving animals. I! 1.
L-
Even when L-carnosine is administered alone to the transplanted area, it exhibits anti-11itl 4 properties, but when used in combination with OK-da32, which is a representative immunostimulant,
-The anti-rejection/cancer properties were enhanced compared to the single administration of carnosine or the single administration of OK-RI32, and the efficacy was particularly marked in cases of disappearance. It also showed effectiveness in measuring liver cancer. The Sarcoma/gO used in 4 is a stock stocked in Komiyama, Kitasato Research Institute for Cancer Control, and other Sarcoma/gO preserved in Japan are also highly proliferative strains, so this one is completely anti-aging. The fact that L-lunosine could be inhibited ensures the antitumor properties of L-lunosine.

As mentioned above, the anti-RMTA effect of lunosine iL-
Cal, Nosine is used to treat various organ cancers and malignant tumors, such as breast cancer, rectal cancer, breast cancer, uterine cancer, oral cavity cancer, esophageal cancer, bile cancer, gallbladder cancer, biliary tract cancer, splenic cancer, kidney liver cancer, and prostate cancer. It can be used alone or in combination for the treatment of cancer, malignant thyroid gland cancer, lung cancer, brain limb cancer, liver cancer, tongue ^W1 thymus gland cancer, skin cancer, volcanic tumor, etc.
By using it in combination with an immunostimulant such as '3.2 to enhance its effect, a very immediate therapeutic effect can be expected.

The anti-inflammatory agent of the present invention may be in any dosage form as long as it is convenient for oral administration and parenteral administration of carnosine for diseases, such as syringes, powders, granules, and tablets. , cassicil agent, enteric-coated agent, ointment, layer agent,
Various dosage forms such as enema and troche are available, and these may be used alone or in combination depending on the patient, the type of liver cancer, symptoms, etc. The clinical dose for adults estimated from basic efficacy experiments is:
Generally, the dose is 0.5 to 31 (oral), and it is preferable to administer the dose in divided doses at appropriate intervals depending on the symptoms.

Since carnosine is easily soluble in water, 3%, 5% and 70% aqueous solutions of carnosine can be easily prepared under aseptic operation. this? The ampoule is injected using a conventional syringe under a stream of inert gas. In addition, the carnosine powder was freeze-dried and sealed in aseptically sealed containers or vials in advance, and immediately before injection, the whole carnosine powder was dissolved in sterile distilled water to give a concentration of 3%, 5% or 19%.
rN may be used for immediate injection as an aqueous solution.

Powders, granules, tablets or capsules for oral administration may be prepared using binders such as syrup, gum arabic, gelatin, sorbitol, tragacanth, forivinyl pyrrolidone, excipients such as lactose, corn starch, calcium phosphate, sorbitol or glycine, Formulated using lubricants such as magnesium stearate, talc, polyethylene golicol, hydroxyglubil methyl cellulose or silica, disintegrants such as potato starch, or fibrillation agents such as sodium lauryl sulfate, in a manner convenient in the art. do. The coating may be performed by any method well known in the tablet ViM industry.

The method for producing ointments is to use finely powdered rat L-carnosine to obtain an ointment of the desired concentration as an ointment base, such as beeswax, spermaceti wax, dehydrated lanolin, etc., according to techniques known in the pharmaceutical industry.
Japanese-French petrolatum, madder grade alcohol, macrogol b or ikegrasuti base (Taisho Pharmaceutical, Ni, similar hydrocar ga ngur ointment base), 11 aqueous ointment listed in the Japanese Pharmacopoeia Law, water-based ointment, or mixtures thereof Mix with this, and add oils such as sesame oil, peanut oil, olive oil, resins, glycerin, propylene glycol, surfactants, bactericides, fungicides, acid L inhibitors, etc. as necessary. Stir thoroughly and knead until homogeneous.

Planing, ointment, and Ik are made in a 4-screw container.For example, 1()1 preservative and L-carnosine are added to the dissolved layer base, mixed uniformly, 1 is poured into a mold, and solidified. Let it out and take it out.

Next, we will discuss formulation examples of the anti-11r1 tumor agent of the present invention.

Formulation Example 1 (injection) Under aseptic operation, 3% of synthesized L-carnosine,
A 1-1 angle was filled with a 3% or 70% (both as L-carnosine) aqueous solution.

Formulation Example 2 (Granules) The following formulation was prepared using synthesized 7tL-carnosine: Carnosine 0.2z Lactose 0.3419- Seven Corn Starch 0. Dasg-Hydroxyglobiglutyl Granules Granules were prepared at 1.00 FI.

Formulation Example 3 (Ointment) Using synthesized carnosine, the following formulation was used as a retardant for hydrocardendal ointment: Carnosine s1 Hydrocardendal IB
An S% soft agent was produced at θg.

Formulation example (small dosage) Fosco S-4,t (Maruuchi Pharmaceutical K.K.) using synthesized lunosine. The following formulation was used as a base (for 1 layer agent) L-carnosine o, a F1 No9 ethyl oxybenzoate 0.00θgsr Phosco S -,5-,5-suitable.

L-carnosine and ethyl hydroxybenzoate total 200
Phosco 5-45 passed through a mesh sieve and dissolved at 30℃
Add it little by little to make it uniform. Addition to the mold was carried out at 3 g°C, and the mixture was left to cool at room temperature in a cooling extractor M cabinet. This was removed from the mold and wrapped in Japanese paraffin paper.

[Brief explanation of the drawing]

FIG. 1 shows the antitumor effect when L-carnosine is used alone, and FIG. 2 shows the antitumor effect when L-carnosine and OK-DA32 are used together. 0 Structural eel ba

Claims (1)

[Claims] An anti-inflammatory drug containing carnosine extract or the like as an active ingredient.