WO1994005321A1 - Moderateur d'activite des enzymes de chassage hepatiques seriques et composition de remede contre les hepatopathies - Google Patents

Moderateur d'activite des enzymes de chassage hepatiques seriques et composition de remede contre les hepatopathies Download PDF

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Publication number
WO1994005321A1
WO1994005321A1 PCT/JP1993/001202 JP9301202W WO9405321A1 WO 1994005321 A1 WO1994005321 A1 WO 1994005321A1 JP 9301202 W JP9301202 W JP 9301202W WO 9405321 A1 WO9405321 A1 WO 9405321A1
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WIPO (PCT)
Prior art keywords
asp
ser
gly
asn
leu
Prior art date
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PCT/JP1993/001202
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English (en)
Japanese (ja)
Inventor
Toshinori Ishizuya
Katsumi Nishimura
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Asahi Kasei Kogyo Kabushiki Kaisha
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.)
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Application filed by Asahi Kasei Kogyo Kabushiki Kaisha filed Critical Asahi Kasei Kogyo Kabushiki Kaisha
Publication of WO1994005321A1 publication Critical patent/WO1994005321A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/225—Calcitonin gene related peptide

Definitions

  • the present invention relates to a novel use of a calcitonin gene-related peptide (Calcin Gene Related Peptide; hereinafter often referred to as "CGRP"). More specifically, the present invention relates to a serum glutamic acid oxa oral transaminase (hereinafter often referred to as "GOT”) and a glutamic acid pyruvate containing a specific CGRP as an active ingredient.
  • GAT serum glutamic acid oxa oral transaminase
  • GTT glutamic acid pyruvate containing a specific CGRP as an active ingredient.
  • the present invention relates to an agent for reducing the activity value of a hepatic escape enzyme such as an acid transaminase (hereinafter often referred to as "GPT").
  • GPT hepatic escape enzyme
  • the present invention relates to a therapeutic composition for a liver disease containing a specific CGRP as an active ingredient.
  • amino acid sequences and peptides are represented using abbreviations adopted by the IUPAC-IUB Biochemical Nomenclature Committee (CNB).
  • CNB Biochemical Nomenclature Committee
  • the liver is the largest single organ in the body, and plays a central role in sugar metabolism, tanbug metabolism, and lipid metabolism.
  • drug metabolism and detoxification are also important functions of the liver, and are therefore vital organs for homeostasis and life support.
  • hepatic drugs are used for liver diseases based on these various causes, but there is still no definitive treatment. This is described in detail in The Journal of Practice Pharmacy, Vol. 41, No. 11. For example, many amino acid infusions and vitamins with specific compositions only have adjunct therapies to supplement essential nutrients, vitamins, etc., which were lacking due to decreased hepatic metabolism. In addition, the effectiveness of antidote such as SH compounds such as daltathione and tiopronin has been questioned.
  • liver diseases such as anti-inflammatory or immunomodulators such as corticosteroids, glycyrrhizin, and azathioprine; protein synthesis promoters such as malotilate; and interferon, which has antiviral activity.
  • Drugs that exert their effects are also known to have serious side effects such as liver dysfunction, hypoaldosteronism, agranulosis, jaundice, and cardiomyopathy.
  • drugs currently used to treat liver disease have shown sufficient efficacy. It did not fully satisfy the clinical situation, for example, because there were concerns about serious side effects.
  • the present inventors first reduced the activity of GOT and GPT, and in some cases, hepatic escape enzymes such as ⁇ -GTP, which are observed in serum as indicators of liver damage.
  • hepatic escape enzymes such as ⁇ -GTP
  • certain CGRPs were surprisingly extremely effective in reducing the levels of GOT and GPT in serum, which are general indicators of liver disease.
  • specific CGRPs enhance their survival rate in acute liver failure models.
  • CGRP has been disclosed in Japanese Unexamined Patent Publication No. Sho 60-501 562 (corresponding to the specification of British Published No. 2 141 433 OA), Japanese Unexamined Patent Publication No. Sho 61-501 101 Gazette (corresponding International Patent Publication W085016658A specification), JP-A-63-258840 (corresponding EP Publication 2730376A specification), etc.
  • Specific CGRP or an acid addition salt thereof obtained by the peptide synthesis method described in (1) has a serum calcium lowering effect (neuropeptides; Vo 1.4, 425 (1984)), a vasodilator effect, and a gastric acid secretion inhibitory effect (Nature Vo 313,54 (198 5), British J.
  • a specific CGRP prevents hepatic cell death due to, for example, carbon tetrachloride and hepatic escape in the culture supernatant which is an indicator of the injury.
  • Certain CGRPs enhance survival in immune-related acute liver failure models, showing markedly reduced enzyme levels.
  • liver damage is indicated by the fact that mature rat hepatocytes are isolated and cultured immediately with hepatotoxic substances such as carbon tetrachloride. At the same time, it is also evident from the fact that, if added in advance, the disorder is remarkably suppressed, and it has been found that the composition is effective for treating liver diseases including the scope of prevention.
  • the present invention has been made based on the above findings.
  • one object of the present invention is to provide a novel agent for lowering the activity value of hepatic escape enzyme, which is an indicator of liver disease and has an increased activity value of hepatic escape enzyme such as GOT and GPT observed in serum. To do that.
  • Another object of the present invention is to provide a method for treating liver disease which is effective.
  • An object of the present invention is to provide an extremely small amount of a novel therapeutic composition for a liver disease.
  • CGRP represented by the formula (I) for decreasing the increased activity of glutamic acid oxamouth acetate transaminase and glutamate pyruvate transaminase in serum.
  • An agent for decreasing the activity of enzyme deviating from serum is provided which contains an effective amount.
  • R is A la — NH—, Ser — NH—, one NH 2 or hydrogen atom
  • Y is one S— or one CH 2 —
  • A is Asp or A sn
  • B is G1y, Asp or G1u
  • C is Leu or Phe
  • D is Va1 or Met
  • E is Va1 or G
  • F is A sn, Ser or A sp
  • G is A sn or Asp
  • I represent Lys or G1u.
  • a therapeutic agent for liver disease comprising an anti-liver disease effective amount of CGRP represented by the formula (I) and a pharmaceutically acceptable carrier, diluent or excipient.
  • a composition is provided.
  • R is a hydrogen atom
  • Y is one S—
  • A is A sn
  • B is Asp
  • C is P he
  • D is Val
  • E is Gly
  • F is A sn
  • G is A sn and I are Lys
  • R is A1a-NH-
  • Y is one S—
  • A is Asp
  • B is Gly
  • C is Leu
  • D is Val
  • E is V
  • al and F are A sn
  • G is A sn and I is Lys.
  • the CGRP represented by the formula (I) as an active ingredient of the agent for lowering the activity of a liver deviating enzyme and a therapeutic agent for liver disease of the present invention also includes an acid addition salt of CGRP represented by the formula (I).
  • the acid addition salts are pharmaceutically acceptable non-toxic salts, for example, salts of inorganic acids such as hydrochloric acid and sulfuric acid, and salts of organic acids such as acetic acid, tartaric acid, succinic acid, and lingic acid. Can be mentioned.
  • the method for producing CGRP represented by the formula (I) is not critical, and it can be produced by various known peptide synthesis methods.
  • Japanese Patent Publication No. Sho 60-501 562 (corresponding to British Published Patent Application No. 2 141 430 A) Japanese Patent Publication No. Sho 61-501 No. 19 (corresponding international patent Published WO 8501658 A), International Patent Publication WO 852804, Japanese Patent Application Laid-Open No. Sho 62-129297, USP 469 Japanese Patent Application Laid-Open No. 7-002, Japanese Patent Application Laid-Open No. Sho 63-3,128,904, Japanese Patent Application Laid-Open No. Sho 63-3-25849, No. ), JP-A-64-26598, JP-A-2-138196, JP-T3-505043, etc. You can refer to the method.
  • the CGRP acid addition salt can be easily produced by allowing an inorganic acid or an organic acid to act on the CGRP product.
  • the activity value of hepatic prolapse enzyme present in the serum of healthy humans is In terms of the unit of measurement (IU), the following is generally used, and the same applies to animals.
  • G ⁇ ⁇ About 5 to about 40 IU / ⁇ ⁇ — GTP About 0 to about 50 IU / ⁇
  • GOT, GPT and ⁇ — GTP activity levels are increased by liver disease, and ⁇ ⁇ ⁇ It is known to increase from ⁇ / ⁇ or more to 300 IU / ⁇ or more.
  • the thus-raised serum hepatic departure enzyme activity value can be effectively reduced by administering an effective amount of CGRP or an acid addition salt thereof.
  • the CGRP or the acid addition salt thereof represented by the above formula (I) has extremely low toxicity, and for example, the compounds (17) and (19) shown in Table 1 as CGRs are intravenously injected into mice. Even if a single dose of 4 mg Z kg (body weight) was administered by each of the subcutaneous routes, subcutaneous route, there were no serious adverse effects or deaths as shown in Table 2 (single dose toxicity test). Not observed. The other compounds shown in Table 1 also showed low toxicity. 5321
  • the CGRP represented by the formula (I) or an acid addition salt thereof may be administered alone, or may be administered as a composition with a pharmaceutically acceptable carrier, diluent or excipient.
  • the component ratio in the composition can be appropriately determined depending on the administration route, administration schedule and the like.
  • CGRP represented by the formula (I) or an acid addition salt thereof may be used in combination with other drugs depending on the condition of the patient.
  • the formulation of the CGRP represented by the formula (I) or an acid addition salt thereof according to the present invention includes, for example, injections, rectal absorbents, vagina Examples include an absorbent, a nasal absorbent, a transdermal absorbent, a pulmonary absorbent, an oral absorbent, an orally administered drug, and the like. These administration forms are not particularly limited, but parenteral administration by injection or the like is preferable. Rectal and vaginal absorbents are generally used in the form of suppositories, nasal absorbents and transdermal absorbents are used in the form of a formulation containing an appropriate absorption enhancer, and pulmonary absorbent is an appropriate dispersant Alternatively, it is used in the form of an aerosol composition containing water and a propellant. Oral absorbents are used in the form of sublingual tablets, for example, with the addition of appropriate absorption enhancers, and oral dosage forms are used in oral forms, such as ribosomes and microcapsules. You.
  • the CGRP of the present invention represented by the formula (I) or an acid addition salt thereof is prepared into a dosage form suitable for each administration form.
  • the carrier, diluent, excipient and the like to be used are not particularly limited, and those generally known can be used. Specific examples will be enumerated below.
  • CGRP represented by the formula (I) or an acid addition salt thereof can be prepared by using a buffer such as acetate buffer or lactate buffer, an isotonic agent such as sodium chloride, or a pH adjusting agent such as hydrochloric acid or acetic acid.
  • the drug is dissolved in distilled water for injection containing an appropriate amount and sterilized by passing through a sterilizing filter to prepare an injection, or CGRP represented by the formula (I) or an acid addition salt thereof is added.
  • Rectal absorbents and vaginal absorbents include CGRP represented by the formula (I) or an acid addition salt thereof, which is a chelating absorption enhancer such as sodium pectinate and sodium alginate; It is prepared as a rectal / vaginal injection suppository or suppository by dissolving or dispersing in distilled water or an oil vehicle by appropriately selecting and using a hypertonic agent such as sodium chloride / glucose.
  • the nasal absorbent is a liquid or powder prepared by adding an absorption enhancer such as dalonic acid, succinic acid or tartaric acid, which is a water-soluble organic acid, to CGRP of the formula (I) or an acid addition salt thereof. It is prepared. Further, a nasal absorbent can be obtained by appropriately adding an emulsion to CGRP represented by the formula (I) or an acid-added salt thereof.
  • an absorption enhancer such as dalonic acid, succinic acid or tartaric acid
  • Percutaneous absorbents can enhance absorption from the skin by adding an absorption enhancer such as Azone (AZone) to CGRP represented by the formula (I) or an acid addition salt thereof. Further, a transdermal absorption agent of CGRP or an acid addition salt thereof represented by the formula (I) can also be obtained by a method using photophoresis.
  • an absorption enhancer such as Azone (AZone)
  • AZone Azone
  • a transdermal absorption agent of CGRP or an acid addition salt thereof represented by the formula (I) can also be obtained by a method using photophoresis.
  • the transpulmonary absorbent is prepared by homogenizing CGRP represented by the formula (I) or an acid addition salt thereof together with a dispersing agent such as Arlacel or Span 80 to obtain a uniform paste. Can be obtained by dispersing the gas in a propellant such as Freon 11 or Freon 12 which has been cooled, and then filling a container equipped with a valve.
  • the oral absorbent may be CGRP represented by the formula (I) or an acid addition salt thereof, for example, ascorbic acids, acidic amino acids, citrates, unsaturated fatty acids, salicylic acids, etc., alone or in combination. Two or more types can be combined and added as a troche, sublingual tablet, powder, etc. by adding an excipient such as glucose and a flavoring agent such as menthol.
  • CGRP represented by the formula (I) or an acid addition salt thereof represented by the formula (I) may be prepared by a method using WZOW emulsion, or may be prepared by a ribosome preparation method.
  • the dosage varies depending on the type of CGRP represented by the formula (I) or the acid addition salt thereof, but it can be generally used in adults in an amount of lg to 200 mg per day, particularly for parenteral administration. In this case, 1 ⁇ g to 10 mg per day is preferred.
  • the number of administrations may be one or two times a day, or one or three times a day or a week.
  • an appropriate amount of CGRP of the formula (I) or an acid addition salt thereof represented by the formula (I) is dissolved in a suitable infusion such as Solita T-3 (manufactured by Shimizu Pharmaceutical Co., Ltd. in Japan), and the solution is intravenously administered for 1 to several hours, for example. It can also be administered intravenously.
  • the amount of CGRP or the acid addition salt thereof represented by the formula (I) in the drug is appropriately determined.
  • the CGRP represented by the formula (I) or an acid addition salt thereof represented by the formula (I) as an active ingredient of the composition for lowering the activity of hepatic deviating enzyme and the therapeutic agent for liver disease of the present invention is prepared by using serum as described in Examples below. Observed during It effectively lowers the elevated activity value of the enzyme that escapes the liver, and at the same time, improves and maintains the impairment of liver function very well.
  • Example 1 Example 1
  • the compound (17) can be obtained according to the synthesis method described in JP-A-63-258490 (corresponding EP Publication No. 270376A).
  • the compound (19) is described in Japanese Patent Publication No. 61-501119 (corresponding International Patent Publication WO8501658A). Those obtained according to the synthesis method were used.
  • Example 2 F344 / DuCrj system rat (body weight: about 200 g) Liver parenchymal cells were isolated by a collagenase perfusion method, and Williams supplemented with 10% fetal serum was added. E medium; suspended in (Wi 11 ms' medium E below "WE medium” intends Rere) were seeded at 5 million cells number Z cm 2 in density di Mesh tissue culture. After 24 hours, replace with WE medium (10% fetal serum added) containing 5. mM carbon tetrachloride, and incubate for another 24 hours. Specified.
  • test plots were prepared in which the test drug was continuously added at various concentrations 4 hours before the addition of carbon tetrachloride and after the addition of carbon tetrachloride.
  • test drugs injections of compound (17) and compound (19) prepared as in Example 1 were used, and 0.2% serum albumin was added for dilution of the compound. Phosphate buffered saline was used. The results are shown in FIGS. 1 and 2.
  • Fig. 1 and Fig. 2 the liver deviating enzyme value (GPT; glutamic pyruvic t ransaminase value, GOT; glutamic oxaloacetic tran saminase value) in the culture supernatant of carbon tetrachloride (CC) was added.
  • GPT glutamic pyruvic t ransaminase value
  • GOT glutamic oxaloacetic tran saminase value
  • CC carbon tetrachloride
  • CGRP represented by the formula (I) directly acts on hepatic parenchymal cells to acquire resistance to hepatotoxicity and promote survival, and this indicates that the formula (I) ) Clearly shows that CGRP is at least one mechanism that can act as a therapeutic agent for liver damage.
  • n indicates the number of mouse models to be tested.
  • An acute liver failure model was prepared in the same manner as in Example 3 with a partial modification of the method of Mizoguchi et al. (Inflammation; Vol. 10, 115-118 (1990)).
  • test drugs injections of compound (17) and compound (19) prepared as in Example 1 were used after diluting them with physiological saline containing 0.2% serum albumin. .
  • Control group 7 1 5 3.8 7.7 Compound (19) administration group 7 5 6 3. 6 4 5.5
  • Figure 1 shows the concentration-dependent effect of compound (17) and compound (19) on the leakage of intracellular enzyme (GPT) in primary cultured rat hepatocytes damaged by carbon tetrachloride (5 mM). It is a Darafu that shows sex.
  • Figure 2 shows the concentration dependence of the inhibitory effect of compound (17) and compound (19) on the leakage of intracellular enzyme (GOT) in primary cultured rat hepatocytes damaged by carbon tetrachloride (5 mM). It is a graph showing the nature. Industrial applicability
  • calcitonin gene-related peptide represented by the formula (I) or the acid addition salt thereof represented by the formula (I) disclosed by the present invention can be used for hepatic such as GOT and GPT in serum as an indicator of liver damage in liver disease. It is a reducing agent of the elevated activity value of the deviating enzyme. Therefore, a pharmaceutical composition containing an effective amount of a calcitonin gene-related peptide represented by the formula (I) or an acid addition salt thereof represented by the formula (I) for an anti-liver disease can be used for drug-induced hepatitis, alcoholic hepatitis, autoimmune disease, It is useful for treating various liver diseases such as inflammatory hepatitis.

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Abstract

L'invention concerne un modérateur d'activité des enzymes de chassage hépatiques sériques qui contient une dose efficace d'un peptide, apparenté à un gène de calcitonine, ou un de ses sels d'addition d'acides, qui permet d'abaisser les activités accrues des enzymes de chassage hépatiques, c'est-à-dire de la transaminase glutamique-oxalo-acétique sérique et de la transaminase glutamique-pyruvique sériques. L'invention concerne aussi une nouvelle composition de remède contre les hépatopathies contenant un ingrédient actif qui comprend le peptide mentionné ci-dessus ou un de ses sels et qui présente des effets secondaires extrêmement réduits. Comme ce modérateur abaisse efficacement les activités enzymatiques accrues qui servent d'indice du degré d'une hépatopathie, on peut utiliser cette composition de remède pour traiter différents types d'hépatopathies, tels que l'hépatite médicamenteuse, l'hépatite alcoolique et l'hépatite chronique active.
PCT/JP1993/001202 1992-09-03 1993-08-26 Moderateur d'activite des enzymes de chassage hepatiques seriques et composition de remede contre les hepatopathies WO1994005321A1 (fr)

Applications Claiming Priority (2)

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JP4/235635 1992-09-03
JP23563592 1992-09-03

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8168592B2 (en) 2005-10-21 2012-05-01 Amgen Inc. CGRP peptide antagonists and conjugates

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0358940A (ja) * 1989-07-27 1991-03-14 Otsuka Pharmaceut Co Ltd カルシウム代謝改善剤
JPH0421699A (ja) * 1990-05-11 1992-01-24 Asahi Chem Ind Co Ltd カルシトニン遺伝子関連ペプチド誘導体

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0358940A (ja) * 1989-07-27 1991-03-14 Otsuka Pharmaceut Co Ltd カルシウム代謝改善剤
JPH0421699A (ja) * 1990-05-11 1992-01-24 Asahi Chem Ind Co Ltd カルシトニン遺伝子関連ペプチド誘導体

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8168592B2 (en) 2005-10-21 2012-05-01 Amgen Inc. CGRP peptide antagonists and conjugates

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