JP4106691B2 - Peptides that inactivate anaphylatoxin C5a - Google Patents

Description

  The present invention relates to a novel peptide that inactivates C5a, which acts as one of the causative substances of inflammatory pathologies including shock, allergy, vasculitis, multiple organ failure, and the like, and a therapeutic drug.

  C5a is a glycoprotein containing 74 amino acids and having a molecular weight of about 11,000. It is generated by being decomposed into C5a and C5b by C4b2aC3b and C3bBbC3b which are C5 converters. It is generated by proteolytic cleavage of the N-terminus of C5, the fifth component of complement (Ntlsson et al., J. Immunol. 114: 815-822 (1975)). The biological properties of C5a extend through numerous cells and tissues involved in both acute and chronic inflammatory processes (Hugli, CRC Crit. Rev. Immunol., 1: 321-366 (1981)). Many of these properties are immunologically beneficial. C5a has been found to mediate host defense mechanisms in response to various pathological conditions. C5a is an inflammatory response, such as smooth microcontraction, increased vascular permeability, occurrence of wheal and redness when injected into human skin, histamine release from mast cells, and oxidation from polymorphonuclear leukocytes (PMNL) Involved in a wide variety of specific biological functions normally associated with the induction of cluster and lysosomal enzyme release. C5a stimulates measurable responses from all circulating white blood cells, including basophils, eosinophils, monocytes and neutrophils (Hugli, supra; Bautsch et al., Immunobiol. 185: 41-52). (1992)).

  Interaction of C5a with polymorphonuclear leukocytes and other target cells and tissues results in increased histamine release, vascular permeability, smooth muscle contraction, and inflammatory cells including neutrophils, eosinophils, and basophils (Hugli, TE, Springer, Semin. Immunopathol., 7: 193-219 (1981)). C5a also plays an important role in mediating the inflammatory effects of phagocytic mononuclear cells that accumulate at sites of chronic inflammation (Allison, A.C. et al., H.U. Agents and Actions, 8:27 (1978)). C5a can induce chemotaxis in monocytes and can cause monocytes to release lysosomal enzymes in a manner similar to the neutrophil response induced by these factors. C5a is known to have an immunomodulatory role by enhancing antibodies, particularly at the site of inflammation (Morgan, E.L. et al., J. Exp. Med., 155: 1412 (1982)).

  Therefore, control of C5a is expected as one of the treatment methods for vasculitis and multiple organ failure caused by a strong inflammatory reaction. Moreover, since C5a may act as an exacerbation factor of asthma and various allergic reactions, control of C5a is expected as one of the means of treatment. Furthermore, since C5a may be involved in various shock symptoms as anaphylatoxin, control of C5a is one of the important treatments in that case. Therefore, the complementary peptide of the present invention having an action of inactivating C5a can be used as a therapeutic agent for various acute diseases including the above-mentioned pathological conditions. For this purpose, C5a receptor inhibitors have been developed as drugs that inhibit C5a, and antibodies against C5a have been developed as those that directly act on C5a and inhibit its action.

  For example, in the invention of JP-T-9-506258, the C5a analog is a C-terminal region of C5a (1-74): N′-Asn (64) -Ile (65) -Ser (66) -His (67 ) -Lys (68) -Asp (69) -Met (70) -Gln (71) -Leu (72) -Gly (73) -Arc (74) -C '(SEQ ID NO: 5) (C5a (1-74 ) Of amino acid sequences 64-) are disclosed.

However, it is not yet possible to control C5a to suppress exacerbation factors of asthma and various allergic reactions, and its appearance is desired.
Japanese National Patent Publication No. 9-506258

  The present invention provides a peptide that directly binds to C5a and inhibits the activity of C5a, and a therapeutic agent for a pathological condition involving C5a.

  In view of the above facts, the present invention uses an amino acid sequence of a peptide consisting of 17 amino acids from the 37th amino acid to the 53rd amino acid, which is considered to play a major role in the activity of C5a, as a target peptide. A peptide having a high complementarity value is designed.

  A feature of the present invention is a peptide that binds to anaphylatoxin C5a and inhibits the activity of C5a, and is a peptide consisting of 17 amino acids whose amino acid sequence is ASGAPAPGPAGPLRPMF when expressed by the single letter code.

  The 37th to 53rd peptide (named PL37) from the amino terminus of C5a acts as an agonist of C5a, and when the peptide is loaded as a MAP peptide, it has the effect of inducing apoptosis in cells with the C5a receptor. I found A complementary peptide to this part (PL37) was designed, and a peptide (named mimetic “A”) having an amino acid sequence of ASGAPAPGPAGPLRPMF when described by the one-letter code of amino acid was obtained. This PepA can bind to C5a and inhibit the activity of C5a.

  Another feature of the present invention is a peptide that binds to anaphylatoxin C5a and has an action of inhibiting the activity of C5a, and is a peptide consisting of 17 amino acids whose amino acid sequence is ASGAPAPGPAGPLRPMF when expressed by the single letter code. The peptide is characterized in that the amino terminal alanine of the peptide is acetylated. Peptide stability can be increased by acetylating the amino terminal alanine of the peptide.

  Another feature of the present invention is that the amino acid sequence has one or two amino acids deleted, substituted, modified, or added, and has an action of inhibiting C5a activity. It is a peptide. A peptide with higher activity and stability can be obtained by chemically modifying the peptide of the present invention or substituting an amino acid. Furthermore, although the peptide of the present invention can be used directly as a drug, a higher-order structure is analyzed using the peptide of the present invention as a prototype peptide, and a chemically synthesized drug having a similar structure is developed based on the knowledge obtained from the analysis. It can also be used.

  Pathological conditions that require inactivation of C5a are assumed to be acute inflammatory conditions, including severe pathological conditions such as anaphylactic shock, vasculitis, multiple organ failure, and DIC (Disseminated Intravascular Coagulation). Yes. Since mimetic “A” (PepA) has an effect of inactivating C5a even in vivo, it can be used as a therapeutic agent for the pathological condition in which such C5a is involved in the pathogenesis.

  In order to save patients in the acute phase of the above pathology, it is necessary to administer a large amount of drug urgently, but peptide agents such as Pep-A are rapidly decomposed in the body, so even if a large amount is administered If it is stopped, it is thought that it will quickly decompose and disappear. Therefore, there are few concerns about side effects such as drug accumulation and sequelae, and this drug can provide a useful means for saving patients in an emergency. Furthermore, it has an effect that can be applied as a prototype for producing a chemically synthesized drug that simulates the higher-order structure of the peptide having a pharmacological effect and mimics the structure.

 Pep-A is rapidly degraded by enzymes in the skin, whereas acetylated A-Pep-A is less susceptible to degradation by the enzymes, and the inhibitory action against C5a continues in the skin. Exhibits inflammatory effects. A-Pep-A is expected as an anti-inflammatory agent.

  Hereinafter, the present invention will be specifically described. The amino acids and single letter notation used in the present invention are shown below. That is, there are 20 kinds of amino acids in nature, and these 20 kinds of amino acids can be divided into hydrophilic amino acids, hydrophobic amino acids and intermediate amino acids according to the structure of the alkyl group. Hydrophilic amino acids include glutamine (Gln; Q), glutamic acid (Glu; E), lysine (Lys; K), asparadine (Asn; N), aspartic acid (Asp; D), tylosin (Tyr; Y) and Histidine (His; H). Hydrophobic amino acids include valine (Val; V), leucine (Leu; L), isoleucine (Ile: I), phenylalanine (Phe; F), and methionine (Met; M). Intermediate amino acids include tryptophan (Trp; W), arginine (Arg; R), glycine (Gly; G), cysteine (Cys; C), serine (Ser; S), proline (Pro; P), alanine ( Ala; A), and threonine (Thr; T).

  The 37th to 53rd peptide (named PL37) from the amino terminus of C5a acts as an agonist of C5a, and when the peptide is loaded as a MAP peptide, it has the effect of inducing apoptosis in cells with the C5a receptor. I found Therefore, a peptide complementary to this part (PL37) was designed, and a peptide (named mimetic “A”) having an amino acid sequence of ASGAPAPGPAGPLRPMF when described by the one-letter amino acid symbol was obtained.

  The peptide thus obtained can be administered to humans in the form of injections, tablets for oral administration, powders and the like. In this case, a preservative, a stabilizer and the like may be included in addition to the above peptide. The above peptides can be treated as appropriate dosage forms after being treated with suitable excipients, adsorption inhibitors and the like used in other biologics. For example, a desired amount of the above-mentioned peptide and preservatives, stabilizers, excipients, adsorption inhibitors, etc. can be mixed as appropriate and filled in an ampule in a certain amount to provide a therapeutic injection. Alternatively, a desired amount of the above peptide may be mixed with a non-antigenic protein component in an aqueous solution, dispensed into vials in a constant amount, and lyophilized to obtain a lyophilized preparation. Such a preparation can be produced in the same manner as the step of producing a normal peptide preparation.

  The following experiment was performed using mimetic “A” (amino acid sequence: ASGAPAPGPAGPLRPMF) and mimetic “B” (amino acid sequence: ASTAPARAGLPRLPKFF) as a control peptide.

  A C5a receptor positive TGW cell line was used as a target cell and examined by the patch clamp method. As a result, when PL37 was bound to MAP (Multiple Antigen Peptide) branched from PL37 with lysine and 8 PL37 were bound to MAP, TGW cells were stimulated, and time was increased. If you leave it, it leads to apoptosis.

  When 500 nM of PL37-MAP is allowed to act on TGW cells, changes in membrane potential can be detected by the patch clamp method as shown in the upper left of FIG. At this time, if 50 micromole mimetic “A” is added to 500 mM PL37-MAP, the membrane potential does not change at all, and the action of PL37MAP is completely suppressed (FIG. 1: upper right). On the other hand, control mimetic “B” does not cause such suppression even when 50 micromole is added (FIG. 1: middle left).

  Clear suppression was also observed at 10 micromolar mimetic “A” (FIG. 1: middle right), but no suppression was observed at 1 micromolar (FIG. 2). The results are summarized in FIG. 2. As shown in FIG. 2, significant inhibitory activity was observed with 50 micromolar mimetic “A”.

Human peripheral blood was collected from a vein, and neutrophils were separated using specific gravity centrifugation or the like.
A fluorescence indicator (Fula2) was incorporated into the separated neutrophils, and the calcium influx measurement that occurred when C5a was allowed to act on the neutrophils was detected with the Hamamatsu Photonics Argus apparatus.

  As a result, when 700 pM of C5a is allowed to act, calcium influx caused by C5a stimulation is detected as a fluorescent signal (FIG. 3: upper left). When mimetic “A” was added to 700 pM C5a with * 700 nM (FIG. 3: upper right) or 350 nM (FIG. 3: second left), the activity of C5a was completely suppressed. Some suppression was also observed at 70 nM and 7 nM (FIG. 3: second row on the right, third row on the left). In contrast, the control peptide mimetic “B” showed no inhibition even at 700 nM and 70 nM. FIG. 4 summarizes the repeated experiments, and a clear inhibitory effect was observed with mimetic “A” peptides of 7 nM or more against 700 pM (0.7 nM) of C5a.

  Male rats weighing approximately 250 g were sensitized with 0.05 mg / kg LPS, and after 20 hours, a monoclonal antibody against Crry (5I2 antibody), a species-specific complement control membrane factor, was administered 0.75 mg / kg intravenously. Causes death within 30 minutes.

All rats escaped from shock death if 4 mg / kg mimetic "A" (abbreviated simply as Pep-A) was administered intravenously 10 minutes before administration of the 5I2 antibody. At 0.5 mg / kg and 2 mg / kg Pep-A, some rats died of shock, but some were able to save lives. In contrast, 4 mg / kg of control mimetic “B” (abbreviated as Pep-B) instead of Pep-A and rats that received only saline were all killed by shock.
(Table 1) LPS (0.05 mg / kg) administered to rats subcutaneously, and 20 hours later, 5I2 antibody (0.75 mg / kg), a species-specific complement control membrane factor, was administered intravenously within 30 minutes Cause death to shock. All patients will survive if mimetic "A" (Pep-A) is administered at 4 mg / kg 10 minutes before 5I2 antibody administration. Control * mimetic "B" (Pep-B) and saline alone cannot be saved.

以上の結果から、mimetic"A"（Pep-A)は生体内でもC5aを不活化してショック死を防ぐ作用を持つことが明らかとなった。

From the above results, it was clarified that mimetic “A” (Pep-A) has the action of inactivating C5a and preventing shock death in vivo.

When CAWS (Candida albicans water solubule extract) derived from Candida albicans is intravenously administered to mice, the mice die by acute inflammation accompanied by complement activation. Prior to this, even if mimetic “A” (Pep-A) was administered 10 minutes before this, it was not possible to save the life. It was thought that Pep-A was rapidly degraded in the mouse body. Therefore, in order to increase the stability of Pep-A to proteolytic enzymes, acetylated Pep-A (A-Pep-A) in which the amino terminal alanine was acetylated was prepared.
(Table 2) In order to investigate the effect of acetylated Pep-A (A-Pep-A) on CAWS lethality, male ICR mice, 5 weeks old were used, and physiological saline or A- CAWS was administered intravenously to ICR mice treated with Pep-A. As a result, suppression was not observed in the system administered with 200 μg CAWS per mouse, but in the system administered with 100 μg CAWS per mouse, lethality was suppressed in the A-Pep-A intravenous (iv) pretreatment group of mice. Was observed. Therefore, the therapeutic effect of Pep-A can be enhanced by chemical modification such as acetylation of amino terminal alanine.

Pep-A and amino for the inflammatory reaction caused by administration of monoclonal antibody (5I2 antibody) against Crry, a complement regulatory membrane factor, into the dorsal skin of rats (Wistar / ST male, 8 weeks old, body weight approximately 250 grams) The action of A-Pep-A acetylated at the terminal alanine was analyzed. As an indicator of inflammatory response, Evans Blue solution (dissolved at a concentration of 8 mg / ml in physiological saline) is injected through 0.5 ml tail vein 1 hour before administration of 5I2 antibody, and the pigment leaks to the inflamed area. (Nishikawa, K., Matsuo, S., Okada, N., Morgan, BP and Okada, HJ Immunol. 156: 1182-1188 (1996)). Since the dye binds to the plasma protein, it is an indicator that the plasma protein has escaped from the capillaries due to the occurrence of a local inflammatory reaction.
(Table 3)
When 5 μg of 5I2 antibody (dissolved in 40 μl of raw food) is inoculated intradermally, 4 μg of A-Pep-A can be mixed to suppress dye leakage after 30 minutes, and 40 μg of A-Pep When -A was added, clear suppression was observed even after 2 hours. On the other hand, in Pep-A, suppression of pigment leakage was not observed even at 30 minutes even when 40 μg was used. It was shown that the inflammation-suppressing action is greatly enhanced by changing Pep-A to A-Pep-A. Pep-A is rapidly degraded by enzymes in the skin, whereas acetylated A-Pep-A is less susceptible to degradation by the enzyme, so the inhibitory action against C5a continues in the skin, causing inflammation. It could be interpreted that the inhibitory action was strong. Therefore, A-Pep-A is considered to exert a strong effect as an inflammation inhibitor.

薬物製剤
治療的有効量の１つまたは複数の識別された上記の作用物質を含む薬学的に許容し得る製剤が、１つまたは複数の薬学的に許容し得る担体（添加剤）および／または希釈剤とともに処方される。以下で詳細に説明するように、本発明の薬学的組成物は、以下のことに適応したものを含めて、固体または液体での投与のために具体的に処方され得る：（１）経口投与、例えば、水薬（水溶液もしくは非水溶液または懸濁液）、錠剤、巨丸剤、粉末薬、顆粒剤、舌に塗布するためのペースト；（２）非経口投与、例えば滅菌溶液もしくは懸濁液として例えば皮下、筋内もしくは静脈内注射による；（３）局所応用、例えば皮膚に応用されるクリーム、軟膏またはスプレーとして；または（４）膣内または直腸内に、例えば膣座薬、クリームまたは発泡剤として。しかし、いくつかの実施形態では、本発明の化合物を単に滅菌水に溶解または懸濁してもよい。

Drug Formulation A pharmaceutically acceptable formulation comprising a therapeutically effective amount of one or more of the above identified agents is one or more pharmaceutically acceptable carriers (additives) and / or dilutions It is prescribed with the drug. As explained in detail below, the pharmaceutical compositions of the present invention may be specifically formulated for administration in solid or liquid, including those adapted to: (1) Oral administration E.g. liquid medicine (aqueous or non-aqueous solution or suspension), tablets, pills, powders, granules, paste for application to the tongue; (2) parenteral administration, e.g. sterile solutions or suspensions For example by subcutaneous, intramuscular or intravenous injection; (3) as a topical application, for example a cream, ointment or spray applied to the skin; or (4) in the vagina or rectum, for example a vaginal suppository, cream or foaming agent. As However, in some embodiments, the compounds of the invention may simply be dissolved or suspended in sterile water.

  The phrase “therapeutically effective amount” as used herein refers to an agent or composition effective to produce any desired therapeutic effect at a reasonable benefit / risk ratio applicable to any medical treatment. It means the amount of things.

  The phrase “pharmaceutically acceptable” is used herein to refer to problems and complications such as excessive toxicity, irritation, allergic reactions, etc., within reasonable medical judgement, commensurate with a reasonable benefit / risk ratio. Without being used to refer to compounds, materials, compositions, and / or dosage forms suitable for use in contact with human and animal tissues.

  The phrase “pharmaceutically acceptable carrier” as used herein refers to carrying or transporting an antagonist of the invention from one organ or part of the body to another organ or part of the body. Means a pharmaceutically acceptable material, composition or excipient, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material involved. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the dosage form and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) Cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) cocoa butter and suppository waxes (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols such as propylene glycol; (11) glycerin, sorbitol, mannitol and polyethylene Polio like glycol (12) Esters such as ethyl oleate and ethyl laurate; (13) Agar; (14) Buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) Alginic acid; (16) Pyrogen-free water; (18) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in drug formulations. In some embodiments, the drug formulation is non-pyrogenic. That is, the patient's body temperature is not increased.

The term “pharmaceutically acceptable salts” in this regard refers to the relatively non-toxic inorganic or organic acid addition salts of the compounds of the present invention. These salts may be prepared in situ during the final isolation and purification of the compounds of the invention, or the purified compounds of the invention can be separated from a suitable organic or inorganic acid in its free base form. And may be prepared by isolating the salt thus formed. Typical salts include hydrobromide, hydrochloride, sulfate, hydrogen sulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, lauric acid Salt, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionic acid Salt, and lauryl sulfonate. (See, for example, Berge et al. (1977), J. Pharm. Sci. 66: 1-19.)
Pharmaceutically acceptable salts of the agents of the present invention include the conventional non-toxic salts or quaternary ammonium salts of the compounds, eg, from non-toxic organic or inorganic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid; and acetic acid, propionic acid, succinic acid, Glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid There are salts prepared from organic acids such as toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isothionic acid and the like.

In other cases, the agents of the present invention may contain one or more acidic functional groups and are thus capable of forming a pharmaceutically acceptable salt with a pharmaceutically acceptable base. In these examples, the term “pharmaceutically acceptable salt” refers to a relatively non-toxic inorganic or organic base addition salt of a compound of the invention. These salts may also be prepared in situ during the final isolation and purification of the agent, or the purified agent in its free acid form, with a pharmaceutically acceptable metal cation. It can also be prepared by reacting separately with a suitable base, such as a hydroxide salt, carbonate or bicarbonate, with ammonia, or with a pharmaceutically acceptable organic primary, secondary or tertiary amine. Good. Typical alkali or alkaline earth salts include lithium salts, sodium salts, potassium salts, calcium salts, magnesium salts, and aluminum salts. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like. (See, eg, Berge et al. (1977), supra).
Wetting agents such as sodium lauryl sulfate and magnesium stearate, emulsifiers and lubricants, and colorants, release agents, coatings, sweeteners, flavors and fragrances, preservatives and antioxidants are also present in the composition. May be.

  Examples of pharmaceutically acceptable antioxidants include: (1) Water soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium hydrogensulfate, sodium disulfite, sodium sulfite and the like; (2 ) Oil-soluble antioxidants such as ascorbyl palmitate, butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol and the like; and (3) citric acid, ethylenediaminetetraacetic acid (EDTA) ), Metal chelating agents such as sorbitol, tartaric acid, phosphoric acid and the like.

  Dosage forms of the present invention include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and / or parenteral administration. The dosage form may be conveniently presented in unit dosage form and may be prepared by any method well known in the pharmaceutical art. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound that produces a therapeutic effect. Generally, out of 100 percent, this amount is the active ingredient ranging from about 1% to about 99%, preferably from about 5% to about 70%, most preferably from about 10% to about 30%.

  The methods of preparing these dosage forms or compositions comprise the step of combining one or more agents of the present invention with a carrier and optionally with one or more accessory ingredients. In general, dosage forms are prepared by uniformly and intimately bringing into association one or more agents of the present invention with liquid carriers or finely divided solid carriers or both and, if necessary, shaping the product.

  Suitable dosage forms of the invention for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (with seasoned active ingredients, usually sucrose and gum arabic or tragacanth), powders, granules, or As a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or pastilles (gelatin and glycerin, or inert groups such as sucrose and gum arabic) And / or a gargle and the like, each containing a predetermined amount of the compound of the present invention as an active ingredient. The agent of the present invention may be administered as a bolus, electuary or paste.

  In solid dosage forms of the invention for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), the active ingredient is one or more pharmaceuticals such as sodium citrate or dicalcium phosphate (1) a filler or bulking agent such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) for example Binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and / or gum arabic; (3) humectants such as glycerol; (4) agar, calcium carbonate, potato or tapioca starch, alginic acid Certain silicates and disintegrants such as sodium carbonate; (6) Absorption enhancers such as quaternary ammonium compounds; (7) Wetting agents such as cetyl alcohol and glycerol monostearate; (8) Absorbents such as kaolin and bentonite clay. (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; and (10) colorants. In the case of capsules, tablets and pills, the drug composition may comprise a buffer. Similar types of solid compositions can also be used as fillers in soft and hard-filled gelatin capsules with excipients such as lactose or lactose and high molecular weight polyethylene glycols and the like.

  A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be binders (eg, gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (eg, sodium glycolate starch or crosslinked sodium carboxymethylcellulose), surfactants Alternatively, it can be prepared using a dispersant. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

  Solid dosage forms such as tablets of the drug composition of the present invention, such as dragees, capsules, pills and granules, are optionally scored or like coatings such as enteric coatings well known in the drug dispensing arts. May be prepared using a unique coating and shell. They use, for example, hydroxypropylmethylcellulose, other polymer matrices, liposomes and / or microspheres in various ratios to provide the desired release profile, and slow or controlled release of the internal active ingredient. May be formulated to provide. They may be sterilized, for example, by filtration through a bacteria retaining filter, or by incorporating a sterilant in the form of a sterile solid composition that can be dissolved in a sterile injectable medium such as sterile water immediately before use. . These compositions may optionally contain opacifiers, in compositions that release one or more active ingredients only in certain parts of the gastrointestinal tract or preferentially there, optionally in a delayed manner. There may be. Examples of embedding compositions that can be used are polymeric substances and waxes. The active ingredient may be in microencapsulated form, if appropriate, with one or more of the above-described excipients.

  Liquid dosage forms for oral administration of the compounds of the invention include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. Liquid dosage forms include, in addition to the active ingredient, inert diluents commonly used in the art, such as water and other solvents, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzoic acid Like fatty acids esters of benzyl, propylene glycol, 1,3-butadiene glycol, oils (especially cottonseed oil, peanut oil, corn oil, embryo oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol and sorbitan Solubilizers and emulsifiers, and mixtures thereof.

  In addition to inert diluents, oral compositions may include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, flavoring and preserving agents.

  Suspensions may be suspended in addition to the active compound, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof. A turbidity agent may be included.

  Dosage forms of the drug composition of the present invention for rectal or vaginal administration may be presented as suppositories. This suppository comprises mixing one or more agents of the present invention with one or more suitable nonirritating excipients or carriers including, for example, cocoa butter, polyethylene glycol, suppository waxes or salicylates. Which is solid at room temperature but liquid at body temperature, it will melt in the rectum or vaginal cavity and release the active compound.

  The dosage forms of the present invention suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray dosage forms containing carriers as known to be suitable in the art.

  Dosage forms for topical or transdermal administration of one or more agents of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active agent may be mixed under sterile conditions with a pharmaceutically acceptable base material and, if necessary, preservatives, buffers, or propellants.

  Ointments, pastes, creams and gels are used in addition to the active compounds according to the invention for animal or vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and zinc oxide. Or an excipient such as a mixture thereof.

  Powders and sprays may contain, in addition to the compounds of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. The spray may further comprise customary high pressure gases such as chlorofluorinated hydrocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

  Transdermal patches have the additional advantage of controlled delivery of the agents of the present invention to the body. Such dosage forms can be made by dissolving or dispensing the compound of the present invention in the proper medium. Absorption enhancers can also be used to increase the flux of the agent of the invention across the skin. The speed of such flux can be controlled by either providing a speed control membrane or by dispersing the compound in a polymer matrix or gel.

It is a figure which shows the change of the membrane potential which made PLGWMAP act on TGW cell. 1 is a drawing summarizing changes in membrane potential caused by the action of PL37MAP on TGW cells. It is drawing which shows the calcium influx which arises by the stimulation of C5a. It is a drawing that summarizes the data of calcium influx caused by C5a stimulation.

Claims (3)

  A peptide that binds to anaphylatoxin C5a and has an action of inhibiting the activity of C5a, the peptide consisting of 17 amino acids whose amino acid sequence is ASGAPAPGPAGPPLRPMF when expressed by the single letter code.   A peptide that binds to anaphylatoxin C5a and has an action of inhibiting the activity of C5a, which is a peptide consisting of 17 amino acids whose amino acid sequence is represented by the one-letter code of amino acid, which is ASGAPAPGPAGPPLRPMF, wherein the alanine at the amino terminal of the peptide is acetyl Peptide characterized by being made. An anti-inflammatory agent comprising the peptide according to claim 1 or 2 , or a pharmaceutically acceptable salt of the peptide.

Images