WO2000021554A1 - Preventives or remedies for cachexia - Google Patents

Abstract

Preventives or remedies for cachexia which contain as the active ingredient osteoclastogenesis inhibitory factor (OCIF) or its variant.

Description

 Description Cachexia preventive or therapeutic agent

 TECHNICAL FIELD The present invention relates to an agent for preventing or treating cachexia comprising an osteoclast formation inhibitory factor (Ost. Eoclastogenesis Inhibitory Factor; hereinafter, referred to as FOCI Fj) as an active ingredient. Background art

 恶: Fluid mainly causes progressive weight loss, anemia, edema, anorexia, etc., which develop in chronic diseases such as malignant tumors, tuberculosis, diabetes, blood diseases, endocrine diseases, infectious diseases and acquired immunodeficiency syndrome. It is a systemic disease that causes symptoms (see J. Parenteral and Enteral Nutrition. Vol. 12, 286-298, (1988): American Journal of Medicine, vol. 85, 289-291. (1988) ).

 In particular, when a patient with a tumor develops cachexia, even if an antitumor agent continues to be administered after the administration of an antitumor agent, side effects such as bone marrow toxicity due to the antitumor agent are added. No improvement in cachexia was observed (see J. Clinical Oncology, vol. 12, 21 3-225. () Ί)).

 In addition, the progression of cachexia, especially in patients with malignant tumors, significantly reduces the patient's physical strength, making it impossible to continue treatment with antitumor drugs, which are generally considered highly toxic. This will hinder the treatment of malignant tumors (see J. Clinical Oncology, vol. 12, 213-225, (1994)).

 In addition, nutritional supplements are often used to improve cachexia symptoms, but such treatments may instead lead to the growth of malignant tumors and shorten patient survival (J Clinical Oncology, vol. 12, 213-225, (1994)).

 Thus, there is a need for the development of a drug that improves the symptoms of cachexia.

On the other hand, OC.1 F is composed of the amino acid sequences described in Amino Acid Numbers 1 to 380 of SEQ ID NO: 1 and has been found as a protein having an activity of inhibiting osteoclast differentiation or maturation. And bone loss such as osteoporosis, osteoarthritis, multiple myeloma, etc. It is known to be useful as a preventive or therapeutic agent for bone metabolism and dysfunction (WO% / 262】 7), however, the protein has a preventive and therapeutic effect on cachexia That isn't true. 'Ϊ;

 The present inventors have shown that OCIF or a mutant thereof exerts an excellent preventive or therapeutic effect on cachexia, and completed the present invention.

 This description is

 (1) A cachexia comprising, as an active ingredient, an osteoclast formation inhibitory factor (OCIF) comprising an amino acid sequence represented by any one of amino acid numbers 1 to 380 of SEQ ID NO: 1 or a mutant thereof. Prophylactic or therapeutic agents,

 () A preventive or therapeutic agent for cachexia comprising, as an active ingredient, an osteoclast formation inhibitory factor (OCIF) comprising an amino acid sequence represented by amino acid numbers 1 to 380 of SEQ ID NO: 1 in the sequence listing,

 About ...

 In the present invention, the term “mutant” refers to a protein in which one or more amino acids in the amino acid sequence of OCIF have been substituted, deleted, added or inserted, and the OCIF activity (inhibition of osteoclast formation) is determined. Activity).

 The OC1 F protein can be produced by the method described in W096 / 26217 ...

 In general, eukaryotic genes are considered to exhibit polymorphism as is known for interferon genes (eg, Nishi, T. et al. (1985) Biochem. HI, 153- 159), this polymorphism may result in the replacement of one or more amino acids, or the nucleotide sequence may be replaced but the amino acids remain the same.

At one or more sites in the amino acid sequence of the OCIF protein consisting of the amino acid sequences of amino acids 1 to 380 of SEQ ID NO: 1, one or more amino acid residues are substituted or deleted. Lost, added or inserted proteins often also have OCIF activity. In the present invention, these proteins are referred to as OCIF mutants. That is, as long as these naturally occurring or artificially synthesized proteins have OCIF activity, all of those proteins can be used in the present invention.

DNA encoding such a protein can be obtained, for example, by the phosphite 'triester method (Nature, ΰ, 105-111, (1984)! [! The codon for amino acid can be selected arbitrarily, for example, considering the codon usage of 使用^: ¾ ·: to be used. (Refer to Nucleic Acid Res. I>, 143-174. (1981).) Further, partial modification of these nucleotide sequence codons can be carried out according to a conventional method. Site-directed mutagenesis / Proc. Natl. Acad. Sci. USA 81, 5662-5666, (] 984) using the primer consisting of the synthetic nucleotide to be used. In addition, any --ό 'or multiple amino acid residues may be deleted. In order to create a mutant, the DNA is trimmed from the end using exonuclease Bal3l (see “Seismological Chemistry Laboratory Lecture 1, Genetic Research Method II”, pages 335-354), cassette mutation (Refer to “Shinogen Chemistry Laboratory Course 2: Nucleic Acid I] I Recombinant DNA Technology”, pp. 242-251.) The codon for the amino acid is known per se, and Selection is optional, and can be determined according to a conventional method, taking into account, for example, the codon usage of the ffj host.

 The OC [F activity can be measured by the method described in W096 / 26217.

 In the present invention, the term "cachexia" refers to a progressive weight loss, anemia, which develops in chronic diseases such as malignant tumors, tuberculosis, diabetes, blood diseases, endocrine diseases, infectious diseases and acquired immunodeficiency syndrome. It refers to a systemic disease with edema and anorexia as the main symptoms. BEST MODE FOR CARRYING OUT THE INVENTION

 Hereinafter, the present invention will be described in more detail with reference to Examples and Preparation Examples, but the present invention is not limited thereto;

Example 1. Anti-cachectic effect of () C 1 P ] A group of 10 Balh / c mice (female, 7 weeks old) were subcutaneously implanted with approximately 2 mm square mouse colon cancer Co Ion 26 cells,

 3 mg / kg, 10 mg / k body weight of OCIF (protein represented by amino acid No. 1-No. 3H0 of SEQ ID NO: 1) produced by the method described in WO 96/26217 After intracellular transfer 7 II, intravenous injection was performed at a rapid rate II. OCIF was 0.0% Tween80 in PBS.

(Phosphaio Ru Her (Hi Saline: lOmM sodium phosphate, 0.15 M sodium chloride, (1) 117.0)). The anti-fluid effect was evaluated using the weight recovery rate calculated by the following formula as an index. Body change (Δ) = body weight on day 12 after cancer transplantation (g) Body weight on day 7 after cancer transplantation

OCTF injection t Weight change of tumor-bearing mice-(Δ _Α ),

Change in body weight of non-OCIF-bearing mice = (Δ _Β ) *,

0 Do not throw-Normal mouse weight change-(A _c ) *,

When

Body ® "yl birefringence index (%) Double (厶_Alpha - delta _beta) / (厶 _{c - Δ Β) X 1 0} 0

* Only the above-mentioned PBS containing 0.01% Tween80 was administered to the mouse without OCIF> -mouse.

Tumor-bearing mice receiving 3 mg / kg of OCIF showed a 31% body weight recovery rate, and tumor-bearing mice receiving 10 mg / kg showed a 50% body weight recovery rate. Example 2, life-span activity of ociF

 Example 1 According to the method described in the above, mouse colon cancer Colon26 cells of about 2 mm square were transplanted under the skin of 10 Balb / c mice (female, 7 weeks old) per group.

 [Example] 30 mg / kg body weight of OCIF (SEQ ID NO: 1) produced by the same method as in

It was intraperitoneally administered twice daily 7 hours after tumor cell transplantation. The life extension activity was evaluated by the survival rate (%) according to the following formula. Life extension (%) '■ (A /-1) X 1 0 0

Λ: 0 C f de throw-Median days of survival in tumor-bearing mouse group

B: 0 C I F non-injected ¥ ft!

 * OCI I ', non-cast mice received only PBS containing 0.01% Tween80. The mice that cast OC1F showed a 100% survival rate. Formulation example 1.

 The cachexia preventive or therapeutic agent containing OC1F of the present invention or a variant thereof as an active ingredient is a water-soluble sterile solution or suspension of adipule dissolved in another pharmacologically acceptable solution. It is also used as a sterile powder formulation (filled in an ampoule with J, preferably a solution of OCIF or a variant thereof, which is preferably dried and dried), and simultaneously diluted with a pharmacologically acceptable solution. Industrial profit ffl potential

 The prophylactic or therapeutic agent for serum of the present invention contains OCIF or a mutant thereof as an active ingredient, and can be administered in various forms. Examples of the administration form include oral administration by tablets, capsules, granules, powders, and syrups, and parenteral administration by injection, infusion, suppository and the like.

 The dosage varies depending on symptoms, age, body weight, etc., but in general, for oral administration, it is about 0.1 mg to 1000 mg per day for an adult, and these may be divided into one or several doses. Can be administered. In addition, for parenteral administration, 0.1 mg or 1000 mg can be administered by subcutaneous injection, intramuscular injection or intravenous injection once.

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Claims

The scope of the claims 1. The cachexia containing an amino acid sequence represented by amino acid numbers 1 to 380 of SEQ ID NO: 1 in the sequence listing, or a cachexia containing an osteoclast formation inhibitory factor or a mutant thereof as an active ingredient乂 is a therapeutic agent. 2. An agent for preventing or treating cachexia, comprising an osteoclast formation inhibitory factor comprising an amino acid sequence represented by amino acid numbers 1 to 380 of SEQ ID NO: 1 as an active ingredient. .