CN101505787B - Treatment of cartilage disorders - Google Patents

Abstract

This invention concerns the treatment of cartilage disorder and osteoarthritis in particular. More specifically, it relates to the use of FGF-18 in treatment regimens and for the manufacture of a medicament for the treatment of patients having a cartilage disorder such as osteoarthritis, such as for example knee osteoarthritis or secondary hip osteoarthritis. Specifically provided is a preferred treatment scheme comprising once weekly administration of an FGF-18 compound per treatment cycle.

Description

The treatment of cartilage disorder

Invention field

The invention belongs to field of medicaments, relate to the treatment of the treatment of cartilage disorder, particularly osteoarthritis.Relate more specifically to the application of FGF-18 in therapeutic scheme and preparing the application for the treatment of on the medicine of suffering from cartilage disorder such as osteoarthritis (for example gonarthritis or Secondary cases coxitis) patient, specifically provide the preferred therapeutic scheme that gives FGF-18 in each treatment cycle once in a week that is included in.

Background of invention

FGF-18 is accredited as a member of FGF family, and it and FGF8 and FGF17 are the most approaching.The activity relevant with FGF18 comprises stimulates Interstitial cell system, particularly myocardial cell, osteoblast and chondrocyte (U.S. Patent No. 6,352,971).FGF18 in conjunction with and activate the IIIc splice variant of FGFR4 and FGFR3 and FGFR2.

Building again of bone is the dynamic process that tissue group and skeletal structure are rely and kept.This process is the equilibrium process between bone resorption and the bone formation, has two class cells to be considered to work therein.These cells are osteoblast and osteoclast.Osteoblast is synthetic to form new bone with deposition substrate.The activity of osteoblast and osteoclast is subjected to a lot of factors (whole body with part) to comprise the adjusting of somatomedin.

Cartilage is a class dense connective tissue, is made up of the cell that is called chondrocyte, is dispersed in the firm gel sample substrate tissue (substrate).Cartilage is avascular (not containing blood vessel), and nutrient is dispersed in the whole substrate.Cartilage sees in joint, rib frame, ear, nose, the throat and between the intervertebral disc.The cartilage that three kinds of main types are arranged: hyaline cartilage, elastic cartilage and fibrous cartilage.The main effectiveness of cartilage provides the skeleton that bone apposition begins.Another significant utility of cartilage is that the activity for articulate bone provides shiny surface and mechanical protection.

The displacement of the damaged cartilage (particularly articular cartilage) that damage or disease cause is a main challenge to the doctor, and effectively treatment is considered to and can not foretells, and only effective in the limited time.In fact the current cartilage injury is effectively treated concentrates on the alleviating pain, and be invalid or nearly unavailable to the regeneration of damaged tissues.Therefore, most of young patient do not seek treatment or are proposed and postpone treatment as far as possible.In the time must treating, standardization program is total joint replacement or microcrack (relating to the subchondral bone puncture stimulates chondrocyte to produce a kind of program of fibrous cartilage deposition).

For the osteoarthritis patient, non-operative treatment comprises that naturopathy, life style change (as taking in sail), bracing, supportive device, oral or injectable drug (as nonsteroid anti-inflammatory drugs) and medical supervision.Surgical operation has specially good effect to the order of severity of osteoarthritis, can make sx, and these symptoms are only short-term activity generally.Tibia or femur otomy (cut skeleton and make wearing and tearing joint reconstruction balance) but mitigation symptoms help to keep positive life style, delay the demand to total joint replacement.Total joint replacement can improve osteoarthritis later stage symptom, but generally needs to change patient's life style and/or level of activation.

Therefore, a kind of method need be arranged in order to treat, to prevent or improve the symptom of cartilage disorder, make damaged tissues regeneration, and this method also need safety and effective as far as possible.In addition, because cartilage disorder can be chronic disease, therefore this method needs and can carry out repetitive therapy to the patient.

Summary of the invention

The invention provides a kind of method for the treatment of the cartilage disorder patient, comprise giving FGF-18 chemical compound, wherein the FGF-18 chemical compound is administered twice at least, and about 4 days of described dosing interval is preferably 5,6,7,8,9 or 10 days.In a preferred embodiment of the present invention, the cartilage disorder for the treatment of is osteoarthritis, the FGF-18 chemical compound is the FGF-18 fragment of being appointed as FGF-18 (170AA) in this description, and the dosiology cycle is weekly intra-articular injection 10-30mcg, continues 3 weeks (treatment cycle).In a preferred implementation, such treatment cycle can repeat after 4 months or 6 months.For example, when adopting the treatment cycle that repeats after 6 months, if first treatment cycle starts from for example certain year January, then second treatment cycle can start from July this year.

The present invention also provides the application of FGF-18 chemical compound on preparation treatment cartilage disorder patient's medicine, and wherein the FGF-18 chemical compound is administered twice at least, and about 4 days of described dosing interval is preferably 5,6,7,8,9 or 10 days.The present invention also provides a kind of FGF-18 compounds for treating cartilage disorder patient's application, and wherein the FGF-18 chemical compound is administered twice at least, and about 4 days of described dosing interval is preferably 5,6,7,8,9 or 10 days.

The present invention also provides the application of FGF-18 chemical compound on preparation treatment cartilage disorder patient's medicine, and described medicine is suitable for being administered twice at least, and described dosing interval is about 4 days, is preferably 5,6,7,8,9 or 10 days.

Detailed Description Of The Invention

The invention provides suitable treatment pattern, comprise with the FGF-18 chemical compound for example FGF-18 (170AA) protein fragments treat the suitable dosage regimen of various cartilage disorders.In the context of the invention, make us finding uncannily that the FGF-18 chemical compound has the best effect of improving disease or symptom to cartilage disorder when the methods and applications administration that discloses according to the present invention.Have found that, when using the dosage identical or suitable with predetermined close of the present invention, the dosage regimen lower than the present invention preset frequency may not be in full force and effect, and the dosage regimen more higher than the present invention preset frequency may be to the result who causes inflammation around cartilage or the joint and/or other is opposite.

Therefore, one aspect of the present invention provides a kind of method for the treatment of the cartilage disorder patient, comprises giving FGF-18 chemical compound, and wherein the FGF-18 chemical compound is administered twice at least, and about 4 days of described dosing interval is preferably 5,6,7,8,9 or 10 days.For example, when dosing interval in the time of about 4 days, administration for the second time can be after the administration first time be carried out in about 96 hours, was when first day morning for example as administration, and the patient will have three days and not accept administration (the 2nd day, the 3rd day, the 4th day), and accept administration again the 5th day morning.

In an especially preferred embodiment, described dosing interval is about 6,7 or 8 days.In a preferred embodiment, their intervals are about 7 days.

In another aspect of the present invention, the application of a kind of FGF-18 chemical compound on preparation treatment cartilage disorder patient's medicine is provided, wherein the FGF-18 chemical compound is administered twice at least, and about 4 days of described dosing interval is preferably 5,6,7,8,9 or 10 days.In an especially preferred embodiment, described dosing interval is about 6,7 or 8 days.In a preferred embodiment, their intervals are about 7 days.

In a preferred embodiment, described administration is each about 7 days at interval.Preferably, the FGF-18 chemical compound is administered once weekly with well-regulated interval.

In a preferred embodiment of the invention, also provide the application of FGF-18 chemical compound on preparation treatment cartilage disorder patient's medicine, described medicine is suitable for being administered twice at least, and described dosing interval is about 4 days, is preferably 5,6,7,8,9 or 10 days.In an especially preferred embodiment, described dosing interval is about 6,7 or 8 days.In a preferred embodiment, their intervals are about 7 days.

In a preferred embodiment, described administration is each about 7 days at interval.Preferably, the FGF-18 chemical compound is administered once weekly with well-regulated interval.

In a preferred embodiment, each treatment cycle gives continuous at least 2 weeks of FGF-18 chemical compound, continuous at least 3 week or continuous at least 4 weeks.In a preferred embodiment, treatment cycle is FGF-18 chemical compound continuous all numbers of administration weekly.In also having a preferred embodiment, each treatment cycle gives continuous 2 weeks of FGF-18 chemical compound, continuous 3 week or continuous 4 weeks, and such treatment can comprise annual 1,2,3,4,5 or 6 treatment cycle.In a preferred embodiment, each treatment cycle gives the FGF-18 chemical compound continuous 3 weeks.In a preferred embodiment, such treatment comprises annual 2 treatment cycle.

In a preferred embodiment, treatment comprises that intraarticular gives FGF-18 chemical compound.Perhaps, treatment comprises that intravenous gives FGF-18 chemical compound.

In further preferred embodiment, treatment comprises that each intraarticular gives FGF-18 chemical compound, and dosage is 1-100mcg, or preferred 1-60 microgram (mcg), or preferred 3-50mcg, or preferred 5-40mcg, or preferred 10-30mcg.In a preferred embodiment, treatment comprises that each intraarticular gives FGF-18 chemical compound, and dosage is about 3,5,10,15,20,25,30,35,40,45,50,55,60mcg.Preferred dosage comprises that each intraarticular gives FGF-18 chemical compound 5,10,15,20,25 and 30mcg.

In further preferred embodiment, treatment comprises that each intravenous gives FGF-18 chemical compound, and dosage is 50-200mcg/kg, preferred 80-120mcg/kg.In a preferred embodiment, treatment comprises that each intravenous gives FGF-18 chemical compound, and dosage is 80,90,100,110 or 120mcg/kg.

In a preferred embodiment, the cartilage disorder for the treatment of with either party's method of the present invention is osteoarthritis, for example is categorized as the osteoarthritis of II level or III level by OARSI.In a preferred embodiment, osteoarthritis can be gonarthritis or coxitis, as the Secondary cases coxitis.Those skilled in the art know employed arthritic classification in this area fully.The OARSI classification is particularly known in the art.Those skilled in the art can be with reference to " Guidelines for the medical management of osteoarthritis " (Marc C.Hochberg, Roy D.Altman, Kenneth D.Brandt, Bruce M.Clark, Paul A.Dieppe, Marie R.Griffin, Roland W.Moskowitz, Thomas J.Schnitzer, Arthritis﹠amp; Rheumatism, the 38th volume, lssue 11,1995. 1535-1546 pages or leaves).

Preferred FGF-18 chemical compound of the present invention is selected from wild type FGF-18 or FGF-18 (170AA).

Chemical compound of the present invention

Natural or wild type FGF-18 is the protein of the chondrocytes expressed of articular cartilage.The present invention relates generally to the application of desmocyte growth factor-21 8 (FGF-18) chemical compound on the treatment osteoarthritis.FGF-18 chemical compound of the present invention comprises for example natural or wild type FGF-18, particularly people FGF-18, its biological activity variant, as the biological activity allele variant, and the biological activity clipped form of FGF-18.The invention still further relates to reservation any variant or the modified forms of bioactive FGF-18 as described in this manual.The activity of the biological activity of FGF-18 described in the following embodiment, the activity on the interior disease model of the special as described body of this description.

The nucleotide sequence of people FGF-18 cDNA is seen SEQ ID NO.1, and the aminoacid sequence of its inference is seen SEQ ID NO.2.FGF-18 was appointed as zFGF-5 originally, in US Patent No. 6,352,971, US5,989,866 and the open communique US2005/0043234 of United States Patent (USP) in have completely and describe, all the elements are all introduced this description as a reference.The analysis of the cDNA (SEQ ID NO.1) of coding people FGF-18 polypeptide has shown that a coding comprises the open reading frame of 207 aminoacid (SEQ ID NO.2) of 180 aminoacid mature polypeptides (207 residues of the 28th residue to the of SEQ ID NO.2).

Corresponding aminoacid sequence shown in mice FGF-18 polynucleotide sequence shown in the SEQ ID NO.3 and the SEQ ID NO.4 be found to have and ethnic group between the high homology of autoploid.At amino acid levels, mice is identical with people's polynucleotide about 98%, has three seed amino acids to change.Those skilled in the art can identify single allele and the polypeptide of SEQID NO.1 or SEQ ID NO.3 and SEQ ID NO.2 or SEQ ID NO.4 difference representative and mice FGF-18 gene, and expection exists allelic variation and alternative splicing.

Just as already mentioned, FGF-18 chemical compound of the present invention comprises for example biological activity clipped form of natural or wild type FGF-18, particularly people FGF-18, its biological activity variant (as active allele variant) and FGF-18.The present invention can relate to any variant or the modified forms of the FGF-18 of the required biological activity of FGF-18 (increasing as the cartilage deposition especially) that keeps this description to describe.

In an embodiment of the invention, the FGF-18 chemical compound is the clipped form of people FGF-18.In a specific implementations, the clipped form of described FGF-18 comprises or is made up of 28-175 residue of SEQ ID NO:2 or functional deriv or variant or mutain (defined as this description).In another embodiment, the clipped form of described FGF-18 comprises or by the 28-176 of SEQ ID NO:2,28-177,28-178,28-179,28-180,28-181,28-182,28-183,28-184,28-185,28-186,28-187,28-188,28-189,28-190,28-191,28-192,28-193,28-194 or 28-195,28-196,28-197,28-198,28-199,28-200,28-201,28-202,28-203,28-204,28-205, a 28-206 or 28-207 residue or functional deriv or variant or mutain (defined as this description) are formed.These polynucleotide, functional deriv or variant or mutain can comprise the-terminal amino acid residue that adds, preferred methionine.Really, depend on expression system and condition, polynucleotide of the present invention can be in recombinant host cell is initial the expression with methionine.

A clipped form that preferred embodiment is FGF-18 of the present invention comprises 170 aminoacid (AA), hereinafter is also referred to as " FGF-18 (170AA) ".Wild type or naturally occurring form are 207 amino acid lengths, and wherein initial 27 aminoacid are signal sequences, and last 11 aminoacid are deleted (also can be confirmed among the naturally occurring FGF-18 in vivo) among the FGF-18 (170AA).FGF-18 (170AA) can be at expression in escherichia coli, owing to there is not signal sequence, the AA sequence is initial with methionine, then is AA28, and end is AA196.The molecular weight of FGF-18 (170AA) is 19.83kDa, pl～10.FGF-18 (170AA) is described as SEQ ID NO.5 hereinafter.FGF-18 (170AA) increases chondrocyte proliferation/differentiation and cartilage deposition, causes the repair and reconstruction of various cartilaginous tissues.

FGF family member's feature is heparin binding domain.Amino acid residue 148 (Gly) to amino acid residue 169 (Gln) zone at SEQ ID NO:2 and SEQ ID NO:4 has identified a FGF-18 heparin binding domain of inferring.By inference, receptor-mediated signal transmission starts from the combination of the FGF part that is compound in the cell surface HSPG.

A lot of FGF family members can put into one or two relevant family according to its 26S Proteasome Structure and Function.AFGF and bFGF comprise three exons being separated by two variable-length introns.FGF-18 comprises 5 exons, and first three is corresponding to first exon of aFGF and bFGF.All known FGF family members engage and form single polypeptide.

FGF-18 ligand-receptor complex analysis has proved that " IIIc " splice variant of the FGFR4 of FGF-18 and FGFR3 and FGFR2 has specificity.The chondrocytes expressed FGFR3-IIIc of cartilaginous tissue and FGFR2-IIIc, these two kinds of receptors see in person joint's cartilage especially.FGFR3 and FGFR2 see mammiferous growth plate, and the formation with the film internal skeleton in cartilage plays an important role.FGFR2 expresses in the mesenchyme of compression at first, and it is beginning that FGFR3 expresses with chondrocyte differentiation and propagation.In skull was grown, FGFR3 saw in cerebral dura mater and the periosteum, and FGFR2 expresses in the osteoprogenitor cell of skeletonization coronal suture.FGFR2 also expresses in trabecular bone.Proved in the past that according to differentiation state and the administering mode of these cells, FGF-18 was that chondrocyte and osteoblastic multiplication agent (are seen United States Patent (USP) 6,352,971 and 5,989,866; Ellsworth et al.Osteoarthritis and Cartilage, 10:308-320,2002; Shimoaka et al., J.Bio.Chem.277 (9) 7493-500,2002).

Be preferably, FGF-18 chemical compound of the present invention increases the cartilage deposition.This increase all can be measured in vivo with in the in vitro tests.Hyaline cartilage, elastic cartilage and fibrocartilaginous propagation are all valuable as therapeutic agent or the component of bio-matrix.The FGF-18 chemical compound, FGF-18 (170AA) for example, and contain FGF-18 compound compositions (" FGF-18 compositions ") and can be used for treating because age related surface fibril forms articular cartilage defect in the synovial joints that causes, the cartilage degeneration that causes because of osteoarthritis and the osteochondral defect that causes because of damage or disease.

FGF-18 chemical compound and compositions also can be used for treating joint disease and the degenerative joint disease that osteochondritis dissecans causes.Rebuilding and field of orthopedic surgery, the FGF-18 compositions also can be used for during tissue defect is rebuild widely self property or expansion of heterologous cartilage and shifts.

FGF-18 chemical compound and compositions also can be used for expanding cell and induce elastic cartilage to generate.Cell expansion and elastic cartilage generate, and to induce regeneration and reparation to ear and nose tissue will be useful.FGF-18 chemical compound and compositions also can be used to expand the chondrocyte population in the culture, are used for self property or heterologous chondrocyte cell transplantation, impose or do not impose the treatment of coexistence then, comprise giving FGF-18 compositions.In these operations, for example, can under arthroscope, gather chondrocyte than the zonule by the unharmed load from impaired joint, chondrocyte is cultivated to increase cell number before the transplanting in the presence of the FGF-18 compositions.Culture with expansion mixes with the FGF-18 compositions then, places articular cavity interior or directly insert damaged place.The FGF-18 compositions can be united use with periosteum or perichondrium graft, and these grafts contain chondrocyte or the fixing cell in place of its precursor that can form cartilage and/or help to implant.The FGF-18 compositions can be united for repairing the cartilage injury with boring under joint lavation, bone marrow stimulation, abrasion arthroplasty, the cartilage or subchondral bone microcrack.In addition, after giving the FGF-18 compositions and making cartilage-derived growth, the treatment that may need to perform surgical operation again makes newly-generated cartilage surface form suitable profile.

By FGF-18 chemical compound of the present invention can also be wild type FGF-18 albumen functional deriv, variant or mutain.

" functional deriv " used herein contained derivant, its variant or mutain and the fusion rotein of FGF-18, and their available methods known in the art are prepared from the functional group that is present in residue side chain or N or C end group group.These functional groups are as long as they remain pharmaceutically acceptable, and namely they do not destroy activity of proteins, are substantially similar to or are better than the activity of FGF-18, and bring toxicity for the compositions that contains FGF-18, namely comprise in the present invention.

These derivants can comprise for example Polyethylene Glycol side chain, and this side chain can improve other activity of protein, for example stability, half-life, bioavailability, human tolerance, or reduce immunogenicity.For reaching this purpose, FGF-18 can be connected in Polyethylene Glycol (PEG), Pegylation can carry out with known method, for example described in the WO 92/13095, can prepare PEG-IFN according to the instruction of WO 99/55377 especially.

Therefore, in a preferred embodiment, the functional deriv of FGF-18 comprises that at least one is attached to the group on one or several functional group, and these functional groups exist with the form of one or several side chain on the amino acid residue.Highly preferred embodiment is that this group is Polyethylene Glycol (PEG).According to the present invention, also can there be several PEG groups to be attached on the FGF-18.

Other derivant comprises modified FGF-18 albumen, for example the long-acting form of FGF-18.The optional especially FGF-18 from Polyethylene Glycolization of long-acting FGF-18, FGF-18-HAS fusion rotein and FGF-18-Fc-fusion rotein.

Other derivant comprises the O-acyl derivative of N-acyl derivative that free amine group and the carboxyl groups (as alkanoyl or carbocyclic ring aroyl) of amide that aliphatic ester, carboxyl and the ammonia of carboxyl or primary amine or secondary amine reaction generates, amino acid residue form or free hydroxyl group (as the hydroxyl of seryl-or Threonyl residue) and carboxyl groups formation.

Used " variant " or " mutain " refers to the analog of FGF-18 in the framework of the present invention, wherein one or several amino acid residue of natural FGF-18 by different radical amino acid replacements the deletion or one or several amino acid residue be added in the natural FGF-18 sequence, the activity of products therefrom is compared to there is no obviously with wild type FGF-18 and is reduced.These mutains are prepared by known synthetic and/or site directed mutagenesis technology or any known technology that other is fit to use.

Comprise as US 5,989 according to " variant " of the present invention or " mutain ", 866 disclose under stringent condition by hybridizing in the DNA of coding FGF-18 or nucleic acid (as DNA or RNA) the encoded protein matter of RNA.Term " stringent condition " refers to hybridize and wash conditions thereafter is the common indication of those skilled in the art " strictness ".See Ausubel et al., Current Protocols in Molecular Biology, Interscience, N.Y., § § 6.3 and 6.4 (1987,1992).The example of stringent condition (nonrestrictive) comprises 12-20 ℃ of the expectation hybridization temperature of wash conditions in being lower than research, and washing is 5 minutes in for example 2 * SSC and 0.5%SDS, and washing is 15 minutes among 2 * SSC and the 0.1%SDS; 0.1 washed 30-60 minute down in 37 ℃ among * SSC and the 0.5%SDS, in 0.1 * SSC and 0.5%SDS, washed 30-60 minute in 68 ℃ then.It will be appreciated by those skilled in the art that stringent condition also depends on DNA sequence, oligonucleotide probe (for example 10-40 base) or the length of mixing oligonucleotide probe.If the use mixed probe is preferably with tetramethyl-ammonium chloride (TMAC) replacement SSC, see Ausbel, the same.

The two or several peptide sequences that the homogeneity reflection is determined by the sequence comparison or the relation between the two or several polynucleotide sequence.Nucleotide pair nucleotide or aminoacid definite when in general, homogeneity refers to that two polynucleotide or two polypeptide are done the comparison of sequence total length respectively are consistent to aminoacid.

When sequence is not definite when consistent, can measure " % homogeneity ".In general, two sequences that compare are alignd, and obtain maximum dependency between these two sequences.This can be included in arbitrary sequence or insert " breach " to increase degree of correction in two sequences.Can measure % homogeneity (what is called is totally compared) to the total length of each sequence that compares, this is particularly suitable for the same or analogous sequence of length, or length short, that determine is measured (so-called local contrast), and this is more suitable for the sequence in different length.

The method of the homogeneity of more two or several sequences and homology and/or approximation is well known in the art.Therefore, for example at Wisconsin Sequence Analysis Package, version 9.1 (Devereux J et al., 1984) program that obtains in, for example BESTFIT and GAP program, can be used for measuring two between the polynucleotide % homogeneity and the % homology between two polynucleotide sequences.BESTFIT uses " local homology " algorithm (1981) of Smith and Waterman, finds the single best region of approximation between two sequences.Other program of homogeneity and/or approximation also is known in the art between the mensuration sequence, BLAST family program (Altschul SF et al for example, 1990, Altschul SF etal, 1997, on the www.ncbi.nlm.nih.gov website, can search from the homepage of NCBI) and FASTA (Pearson WR, 1990).

According to " variant " of the present invention or " mutain ", comprise the protein that fully copies aminoacid sequence with FGF-18 sequence, for example have the protein to the similar substantially activity of FGF-18.

In a preferred embodiment, any such variant or mutain have and FGF-18 sequence 40% homogeneity or homology at least, be more preferred from least 50%, at least 60%, at least 70%, at least 80%, or the best is at least 90% homogeneity or homology.

Can be used for the mutain of FGF-18 of the present invention or its coding nucleotide and comprise the finite set sequence that conforms to substantially peptide or polynucleotide as an alternative, they can need not too much experiment and acquisition routinely by instruction and the guide of those skilled in the art by this description.

Preferential variation according to mutain of the present invention is the variation that is called " guarding " displacement.The conservative aminoacid substitutions of FGF-18 polypeptide can comprise the synonym aminoacid in the group, and they have enough similar physicochemical property, and the displacement between this group membership will keep the biological function (Grantham, 1974) of molecule.Obviously, also can insert and delete in the sequence defined above, only relate to a small amount of aminoacid if particularly insert or delete, for example below 30, be more preferred from below 10, and do not remove or replace aminoacid such as cysteine residues to the functional conformation key.Protein and mutain that such deletion and/or insertion produce fall into scope of the present invention.

Preferably, simultaneous administration is that Table A limits.More preferably, simultaneous administration is that table B limits.Most preferably, simultaneous administration is that table C limits.

The preferred simultaneous administration of Table A

Aminoacid synonym group

Ser Ser，Thr，Gly，Asn

Arg Arg，Gln，Lys，Glu，His

Leu Ile，Phe，Tyr，Met，Val，Leu

Pro Gly，Ala，Thr，Pro

Thr Pro，Ser，Ala，Gly，His，Gln，Thr

Ala Gly，Thr，Pro，Ala

Val Met，Tyr，Phe，Ile，Leu，Val

Gly Ala，Thr，Pro，Ser，Gly

Ile Met，Tyr，Phe，Val，Leu，Ile

Phe Trp，Met，Tyr，Ile，Val，Leu，Phe

Tyr Trp，Met，Phe，Ile，Val，Leu，Tyr

Cys Ser，Thr，Cys

His Glu，Lys，Gln，Thr，Arg，His

Gln Glu，Lys，Asn，His，Thr，Arg，Gln

Asn Gln，Asp，Ser，Asn

Lys Glu，Gln，His，Arg，Lys

Asp Glu，Asn，Asp

Glu Asp，Lys，Asn，Gln，His，Arg，Glu

Met Phe，Ile，Val，Leu，Met

Trp Trp

The preferred simultaneous administration of table B

Aminoacid synonym group

Ser Ser

Arg His，Lys，Arg

Leu Leu，Ile，Phe，Met

Pro Ala，Pro

Thr Thr

Ala Pro，Ala

Val Val，Met，Ile

Gly Gly

Ile Ile，Met，Phe，Val，Leu

Phe Met，Tyr，Ile，Leu，Phe

Tyr Phe，Tyr

Cys Cys，Ser

His His，Gln，Arg

Gln Glu，Gln，His

Asn Asp，Asn

Lys Lys，Arg

Asp Asp，Asn

Glu Glu，Gln

Met Met，Phe，Ile，Val，Leu

Trp Trp

The most preferred simultaneous administration of table C

Aminoacid synonym group

Ser Ser

Arg Arg

Leu Leu，Ile，Met

Pro Pro

Thr Thr

Ala Ala

Val Val

Gly Gly

Ile Ile，Met，Leu

Phe Phe

Tyr Tyr

Cys Cys，Ser

His His

Gln Gln

Asn Asn

Lys Lys

Asp Asp

Glu Glu

Met Met，Ile，Leu

Trp Met

The example that can be used for obtaining producing in the protein of the used FGF-18 mutain of the present invention amino acid replacement comprises any known method step, and for example following United States Patent (USP) proposes: authorize the No of Mark etc., 4,959,314,4,588,585 and 4,737,462; Authorize the No.5 of Koths etc., 116,943; Authorize the No.4 of Namen etc., 965,195; Authorize the No.4 of Chong etc., 879,111; With the No.5 that authorizes Lee etc., 017,691; And the lysine in the U.S. Patent No. 4,904,584 (Shaw etc.) replaces albumen.

The preparation of FGF-18 chemical compound and administration

The FGF-18 chemical compound can be mixed with pharmaceutical composition, namely with preparations such as pharmaceutically acceptable carrier, excipient." pharmaceutically acceptable " is defined as and how comprises carrier, and they are the bioactive effectiveness of interferon activity composition not, and to accepting host's avirulence of administration.For example, for parenteral, activated protein can be mixed with unit dosage forms in saline, glucose solution, serum albumin and Ringer ' s liquid.The preparation of such FGF-18 chemical compound also comprises at least a pharmaceutically acceptable carrier, excipient etc., is called " FGF-18 compositions " in this manual.

FGF-18 chemical compound and compositions can be injected directly in the synovial membrane liquid in joint, or be injected directly into damaged place, can use separately or mix the back use with the carrier that be used for to prolong the suitable carrier that protein discharges (for example suitable slow releasing preparation excipient, as cyclodextrin) or partial restriction release (for example by biocompatibility sponge, similar delivering drugs such as bio-matrix, coated cell).

The preparation that vein (IA) is used should meet the most of requirements that also are applicable to other ejection preparation, namely they need sterilize and be suitable for the injection site (as knee joint, physiological condition SF).The available autoclaving of the sterilization of solution (if all the components of preparation is enough to tolerate thermal stress) or filtration sterilization method reach, and for other preparation, for the preparation method of guaranteeing to obtain the required usefulness of sterile product can be more compound.For example, can not carry out filtration sterilization for the preparation that contains granule (suspensoid), semisolid or solid preparation.In order to make preparation be suitable for the physiological condition of injection site, must consider the feature of SF.For this reason, suitable preparation of the present invention etc. oozes.The pH of preparation is near the pH (being pH7.4) of SF or lower slightly, but suitable be to be not less than pH～5.5 so that chemical constituent reaches the suitableeest stability, but not the issuable side effect of physiology pH value such as activator protein hydrolytic enzyme (as cathepsin) then drop to minimum.The excipient that is used for the IA injection also can be present in other ejection preparation (as muscle or the subcutaneous preparation of using).

In one embodiment of the invention, the administering mode of FGF-18 described herein is selected from following group: administration and intra-articular administration around administration in administration in administration in administration in ear administration, auricle week administration, intranasal administration, nose week administration, the nasal sinuses, the rib, rib week administration, the thorax, thorax week administration, epidural administration, vertebra week administration, synovial membrane week administration, synovial membrane intracavitary administration, the nasal sinuses, the joint.In a preferred embodiment, FGF-18 chemical compound described herein adopts (around the joint administration) or intraarticular (at intra-articular administration) administration around the joint.In one embodiment of the invention, around the joint and intra-articular administration be to be selected from hip joint, knee joint, elbow joint, carpal joint, ankle joint, spinal column, joints of foot, articulations digitorum manus, toe joint, joints of hand, shoulder joint, rib joint, scapula, thigh, tibia, heel and along carrying out in the joint at the spinous process place of spinal column.In a further preferred embodiment, around the joint and intra-articular administration be to carry out at hip joint or knee joint peripheral or intraarticular.

Cartilage disorder

The present invention relates to treat, prevent or improve the method for cartilage in mammals obstacle.Preferably, such cartilage disorder derives from the damage of traumatic injury or osteomalacia.Be appreciated that the people is advisable with the patient who treats by the present invention.Yet, be appreciated that the available method of the present invention of other mammal (including but not limited to Canis familiaris L., horse etc.) treats.

The described method treatment of available description, prevention or the cartilage disorder that improves include but not limited to arthritis, osteochondritis, costal chondritis (as painful nonsuppurative swelling of costal cartilage (Tietze ' s syndrome)), osteomyelitis, polychondritis, relapsing polychondritis and osteochondritis dissecans.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is arthritis.Be preferably, described disease be selected from ankylosing spondylitis, idiopathic diffuse hyperostosis (DISH), gout, pseudogout, infective arthritis, osteoarthritis, rheumatoid arthritis, arthritic psoriasis, reactive arthritis, scleroderma,

Syndrome and Still ' s disease.In a preferred embodiment, the cartilage disorder for the treatment of, preventing or improve is rheumatoid arthritis or osteoarthritis.In an especially preferred embodiment, the cartilage disorder for the treatment of, preventing or improve is osteoarthritis.

Arthritis relates to damage and the corresponding inflammatory process of the articulation structure (joint) of health.Arthritis comprises osteoarthritis, rheumatoid arthritis, conduct damage result's arthritis joint, etc., be common inflammation, they benefit from the therapeutic use of FGF-18 of the present invention.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is ankylosing spondylitis.Ankylosing spondylitis (AS) is the arthritis of involving spinal column.It causes back pain and stiff and crooked, and this is the result of joint of vertebral column (vertebra) carrying out property swelling and stimulation.The patient with severe symptoms, the inflammation of vertebra finally can cause merging, and causes serious limitation of activity.The inflammation that the tendon that connects the joint and the joint is provided support and ligament take place can cause rib, scapula, hipbone, femur, tibia, heel and along pain and the tenderness of the spinous process of spinal column.

Ankylosing spondylitis is a kind of chronic inflammatory arthritis, and it influences joint of vertebral column.The flag sign of AS is to involve joint-sacrum ilium (SI) joint of (spinal column connects pelvis at this place), spinal column bottom.

The course of disease has very big variation, and some individuality is accidental of short duration back pain only, and other individuality can have chronicer serious back pain, As time goes on causes spinal column in various degree stiff.On nearly all case, this sick feature is the acute pain outbreak and alleviates (cycle that problem solves).

Over the years, AS is called with a lot of titles, comprises poker back, rheumatoid spondylitis and ankylosing spondylitis.Since the beginning of the seventies, along with about the increasing of this sick knowledge, an almost term-ankylosing spondylitis of widespread usage has been arranged.AS is a member of attacking in gang's disease of spinal column.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is idiopathic diffuse hyperostosis.The feature of idiopathic diffuse hyperostosis (DISH) is that the vertebra side along spinal column has excessive osteogenesis.It also relates to tendon is connected bone with ligament place (for example ancon, knee and heel) and is inflamed and osteogenesis.The philtrum of suffering from DISH often has bony spur.

DISH (being called Forestier ' s disease sometimes) is considered to a kind of degenerative osteoarthritis, and it is characterized by along the vertebra side of spinal column has excessive osteogenesis.It also relates in tendon is connected the health of bone with ligament other zone (for example ancon, knee and heel) and being inflamed and calcification (osteogenesis).These inflammation and osteogenesis can cause bony spur, and for example the heel spur is common in DISH crowd.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is gout.Gout is a class arthritis, and raise surpasses normal level to uric acid in this disease (a kind of natural refuse that is present in the body).It is removed from kidney by urine unlike usually, but forms crystallization deposition in the joint.These deposits cause arthritis, produce pain, swelling, rubescent and tenderness.The most typical joint of getting involved is hallux toe, but gout also can influence ankle, knee joint, foot, hands, wrist and elbow.Uric acid crystal also can be in other zone as forming deposition under the skin or in other soft tissue and kidney or the urinary tract.

Gout typically influences the joint of hallux toe root.In surpassing the initial onset of half, this is first joint of getting involved.Almost any other joint all can be involved, but gets involved than upper limb joint in the joint of more common lower limb.

The initial onset of most of gouts only relates to a joint, disappears in three to ten days in therapeutic process.Can recurrence in the man-year of more than half gout acute attacks.As time goes on, it is more frequent that outbreak may become, and the persistent period is longer, often involves a plurality of joints.

For the people that some outbreaks delay, disease becomes chronic.The uric acid crystal that is deposited on intraarticular and surrounding soft tissue causes the destructive change of intraarticular, produces the inflammation that continues.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is pseudogout.Pseudogout is the class arthritis that the calcium accumulation causes in the body.Pseudogout results from the accumulation of intraarticular calcium crystallization (calcium pyrophosphate dihydrate).Calcium forms the intraarticular of crystallization deposition between bone, produces this regional pain.Calcium deposition and chronic inflammatory disease can cause that the part articulation structure dies down and damages.

May begin in the pseudogout cartilage to break and produce the hole, in the joint, produce more pain and swelling.As time goes on, cartilage may wear and tear fully, and bone rubs.

As cartilage injury's result, a lot of pain of pseudogout are to help the muscle of joint motion and other tissue (as tendon and ligament) to be forced to the result who causes with unplanned mode work done.Itself does not have neurocyte cartilage, therefore can not feels pain, but muscle, tendon, ligament and influential point of bone feels pain.After cartilage corroded for many years, bone rubbed in fact together.Bone further aggravates the pain to this friction of bone.Bone also can thicken and form lump, is called spur or hyperosteogeny, and they rub together.

As goat, pseudogout can take place suddenly at an intraarticular, the pain of showing effect repeatedly and swelling.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is infective arthritis.Infective arthritis causes arthralgia and swelling.Inflammation is caused that by microorganism microorganism can be antibacterial, virus or fungus.Usually have only a joint involvement, but sometimes two or three joint involvement can be arranged.If the early treatment does not continue usually for a long time.

Infective arthritis infects big joint (shoulder, hip, knee joint) at most, but less joint (finger, ankle joint) also can involve.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is osteoarthritis.Osteoarthritis is arthritic most common form, can be damaged by cartilage to cause.Cartilage overwhelms broken, causes arthralgia and swelling between the bone.As time goes on, cartilage can wear and tear fully, and bone can the phase mutual friction.

Osteoarthritis can influence any joint, but is usually directed to hands and weight-bearing joint such as hip joint, knee joint, foot and spinal column.

The condition of illness of osteoarthritis (OA) is complicated, polyfactorial PD, and it is non-inflammatory in essence, is feature with the general intraarticular articular cartilage degeneration relevant with the age.The feature that OA also has is that the chondrocyte activation causes cell proliferation and apoptosis, protease to be expressed and unusual substrate generates, and the repair of cartilage of decline causes the disappearance of extracellular matrix, substrate calcification and chondrocyte to generate.The degeneration of cartilage and extracellular matrix structure cause bone and the nerve in the joint of getting involved between friction increase.OA makes the pain that is subjected to these sick people that torment to produce various degree unable with carrying out property.Treatment for OA is palliative treatment or surgical intervention at present.

In healthy joint, cartilage plays the amortisseur effect when gravity is added on this joint.The shiny surface of cartilage moves skeleton smoothly.When knuckle shaped becomes osteoarthritis, cartilage roughening attenuation gradually, following skeleton thickening.

Though the usual earlier of disease does not have swelling, along with arthritic progress, may have inflammation.A small amount of cartilage may fracture, and swims in intraarticular, and this has upset other soft tissue of intraarticular, can cause pain and swelling between skeleton.

As time goes on, the cartilage wearing and tearing, skeleton can form lump in its end.These lumps are called spur.Perhaps, cartilage can wear and tear fully, and skeleton can directly be in contact with one another.

OA can cause other problem, and for example: the muscle of fixing joint is useless to be weakened with property, and As time goes on, joint shape is lax, can not work at all.OA influences weight-bearing joint usually, as hip, knee joint, foot and spinal column.But also can involve as non-bearing joints such as articulations digitorum manus and thumb root joints.It does not influence other joint usually, unless when they are injured or be under the ABNORMAL STRESS.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is rheumatoid arthritis.Rheumatoid arthritis (RA) is a kind of chronic autoimmune inflammation disease, mainly involves the multi-joint Synovial membrane, and the result is articular cartilage damage.Pathogeny is that the T lymphocyte relies on, and with rheumatoid factor, at the generation of the autoantibody of self IgG, consequently the formation of immune complex reaches high level in joint fluid and the blood.These complex in the joint can cause that lymphocyte and mononuclear cell soak in the synovial membrane significantly, significant synovial fluid takes place then change; Adding various neutrophilic leukocytees makes articular cavity/joint fluid by similar cellular infiltration.Getting involved, to organize mainly be the joint, and usually be symmetric form.The enzyme that inflammatory cell discharges can digest skeleton and cartilage.As the result of rheumatoid arthritis, skeleton and cartilage can be attacked and damage to (synovial membrane) in the inflamed joints village, causes joint damage, serious pain and other physiological effect.Affected joint can lose its shape and arrangement, causes pain and motor capacity forfeiture.Therefore, RA can cause handicap and improve mortality rate.

Joint injury even can come across the not serious situation of pain, even can take place in early days in disease.The people who much the suffers from RA visible damage on 2 years inherent handss, the sufficient X-line sheet of falling ill.But when X-line sheet is pinpointed the problems, just treat in the time of to be too late.

Serious damage can cause permanent joint deformity and deformity.Pain and swelling can cause difficulty in walking.

The various cytokines of local generation in the joint of Rheumatoid.Studies have shown that much two kinds of prototype proinflammatory cytokine IL-1 play significant feature with TNF-α in the mechanism relevant with the damage of carrying out property joint with synovial membrane inflammation.Really, TNF-α and IL-1 inhibitor can produce the improvement of the clinical and physiology's sign of noticeable inflammation and skeleton corrodes and the radiology sign of cartilage damage is reduced to come into operation for RA patient.But, although these challenging results are arranged, also have the patient of suitable percentage ratio to these two kinds of material Fails To Responds, point out also to relate to other medium in the arthritic pathophysiology.

Difference RA and the arthritic important method of other type are the patterns according to the joint of getting involved.For example, RA influences wrist and a lot of joints of hand, but does not influence the joint of distal-most end usually.

Among the RA, the tendency that the joint has symmetric form to get involved, if the i.e. articulations digitorum manus inflammation of the right hand, the articulations digitorum manus of left hand also may inflammation.Other joint that RA involves is elbow, shoulder, neck, jaw, foot, ankle, knee joint and hip joint normally.Except cervical region, spinal column is not directly got involved in RA usually.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is psoriatic arthritis.Psoriatic arthritis is a kind of condition of illness that produces swelling and pain around intraarticular and joint.It also causes the squamous erythra at skin.

It can influence a lot of joints, comprises finger, wrist, knee joint, ankle, elbow and shoulder joint, spinal column and lower back joint (being called sacroiliac joint).

Psoriatic arthritis also influences the joint surrounding tissue, comprises tendon and ligament.It can cause whole finger swelling, is called " sausage " and refers to or toe.Also has scytitis, particularly on ancon, knee and scalp.Psoriatic arthritis is associated with psoriasis, and psoriasis is to cause a skin inflammation and cover a kind of disease of silver color or Lycoperdon polymorphum Vitt squama.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is reactive arthritis.Reactive arthritis relates to the arthralgia that was caused by former infection, tetanic or swelling.It is most commonly in joint of the lower extremity (knee joint, ankle, toe), but also can appear at upper limb.Problem may be only in the joint or relate to other body system, as eyes, skin, muscle or tendon.When its abarticular zone of invasion and attack, reactive arthritis is called Reiter ' s syndrome.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is scleroderma.Scleroderma is the disease of a kind of pachyderma and hardening.Scleroderma has two types.A kind of is local scleroderma, mainly attacks skin, also can involve muscle and joint.Another kind is systemic scleroderma, and invasion and attack skin and internal organs are as heart, lung and kidney.

Sclerodermatous maximum feature is the hard cicatrix sample fibrous tissue of accumulation in the skin.More unconspicuous variation comprises the damage that little cells of vascular wall is, and then can damage main organs.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is

Syndrome.

Syndrome is salivary gland to be produced damage cause xerostomia, lachrymal gland is produced a kind of chronic disease that damage causes xerophthalmia.It also can attack other position of health, comprises joint, muscle and nerve, such as organs such as lung, kidney, liver, pancreas, harmonization of the stomach brains, such as bodies of gland such as thyroid.

Syndrome can cause these zones any one destroy fully.Because

Syndrome can influence liver and pancreas, so big chance generation lymphoid tissue cancer is arranged.But this is the rare result of extraordinary.

Syndrome can exist in two ways.When it comes across the philtrum of rheumatically or connective tissue disease such as lupus, scleroderma or polymyositis, be " Secondary cases "

Syndrome.When xerophthalmia and xerostomia during without rheumatism, it is called " constitutional "

Syndrome.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is Still ' s disease.Still ' s disease be a kind of be the arthritis of feature with high spiking fever, salmon color erythra and arthritis.This disease is most commonly in the child, often relates to the special sex infantilism arthritis of sending out of whole body.Still ' s disease also is found in the adult, though common like that not as good as the child.This situation is called Still ' s disease or the AOSD of the morbidity of growing up.

As discussed above, the cartilage disorder for the treatment of, preventing or improve described herein comprises: stripping off property chondritis, costal chondritis (as painful nonsuppurative swelling of costal cartilage), osteomyelitis and relapsing polychondritis.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is osteochondritis dissecans.Osteochondritis dissecans (OCD) is a term about osteochondral fracture.Bone cartilage fragment can be present in the original place, not exclusively separates or separate fully.OCD is a kind of osteochondrosis that is limited to the joint epiphysis.The joint epiphysis fails because of compression, and its pathology may relate to wound and ischemia.Wound is primary injury most likely, and ischemia then is secondary injury.

Wound can by direct wound such as impaction fracture or repeatedly Wicresoft hinder as extraordinary compressive stress causes.

Knee joint is the most normal position of getting involved, and elbow joint, ankle joint, tarsal navicular, hip joint, shoulder joint, glenoid cavity, carpal joint also can be got involved.

OCD tends to influence young patient.Among the elbow joint OCD, average 23 years old (4-47 year) of patient age.Among the ankle joint OCD, average 20 years old (8-50 year) of patient age.Among the hip joint OCD, average 24 years old (14-39 year) of patient age.

The patient often tells at range of movement extremely pain.Heating sense when joint peripheral edema and palpation are arranged usually.When lower limb were got involved, the patient often had limping.Joint protectiveness braking can improve symptom usually.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is costal chondritis.Costal chondritis is the inflammation of the node between last rib and the cartilage that is fixed in breastbone (breastbone or sternum), and reason is not bright.When the pain of costal chondritis during with swelling, be called Tietze ' s syndrome.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is osteomyelitis.Osteomyelitis is a kind of bone of suppuration bacterium infection and acute or chronic inflammation process of structure thereof of being secondary to.

But osteomyelitis with infection localization maybe can send out whole periosteum, cortex, medullary substance and cancellous tissue.Bacterial pathogen is different with infection mechanism because of patient age.

Hematogenous osteomyelitis is by the infection of antibacterial due to the blood dissemination.Skeleton actute infection due to acute hematogenous osteomyelitis is sent out in bone from remote origin with antibacterial is feature.This disease mainly betides the child.Common site is the skeleton in the growth of growth fast and the metamorphosis of blood vessel height.When blood vessel when distal metaphysis forms acute angle, blood flow obviously slowly or deposition makes blood vessel easily form thrombosis, the local necrosis of skeleton own and antibacterial are sent out.Acute hematogenous osteomyelitis (although title is like this) can have chronic clinical development and insidious onset.

Directly or in abutting connection with the osteomyelitis of inoculation during by wound or surgical operation tissue and antibacterial directly contact cause.Direct inoculation (in abutting connection with focus) osteomyelitis be secondary to organism from the inoculation of direct site of injury, from the adjacency focus of infection send out or surgical operation after septicemia.The myelitic clinical manifestation of direct inoculation easily relates to multiple organism than hematogenous osteomyelitis localization comparatively.

Other kind comprises chronic osteomyelitis and is secondary to the osteomyelitis of peripheral blood vessel.No matter the initial cause of disease and/or the mechanism of chronic osteomyelitis, no matter have or not powerful the intervention, it all is to continue and recurrent.Though peripheral blood vessel is classified initial reason as, it is actually the true cause of a kind of susceptible factor rather than infection.

Make patient easily suffer from myelitic morbid state and comprise diabetes, sickle cell disease, acquired immune deficiency syndrome (AIDS) (AIDS), intravenous drug abuse, alcoholism, life-time service steroid, immunosuppressant, chronic joint disease.In addition, the prothesis orthopedic device is a kind of independently risk factor, and recent orthopaedic surgery or open fracture also are like this.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is relapsing polychondritis.Relapsing polychondritis (RP) is the rare and serious ictal inflammation of involving cartilage structure, mainly involves ear, nose and the tree-shaped structure of larynx tracheal bronchus.Other structure of getting involved can comprise eyes, cardiovascular system, tip joint, middle ear and internal ear.

Etiology the unknown that this is sick, but pathogeny likes the autoimmune problem in nature very much.The pathology that the evidence of the autoimmune cause of disease comprises that its clinical manifestation is relevant with other autoimmune disease, invade profit CD4 T cell and antigen antibody complex in haploid related, the cartilage of getting involved of itself and HLA-DR4 find, to cell and the humoral response of collagen I I type and other collagen antigen and observe the common optimal inhibition of immunosuppressant therapy scheme should disease.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is the cartilage injury due to the wound.The cartilage injury can be used as the result of traumatic mechanical damage and occurs.Directly strike or other wound can be damaged cartilage.Cartilage does not have direct blood supply, therefore almost can not self repair.Method of the present invention is promoted repair of cartilage, and in one embodiment of the invention, the cartilage injury that wound causes results from thunder bolt or operation.In a special embodiment of the present invention, the cartilage injury that wound causes results from operation, particularly orthopedics or plastic surgery operations.The present invention also considers the treatment with move relevant damage or the relevant joint tissue wearing and tearing with motion.

In one embodiment of the invention, the cartilage disorder for the treatment of, preventing or improve is the outer surface diseases of non-beauty treatment.In such embodiments, can use method of the present invention and the purposes that is associated with the plastic surgery.

After describing the present invention fully, skilled person in the art will appreciate that the inventive method can carry out and do not deviate from the spirit and scope of the present invention in the scope of very wide equivalent parameters, concentration and condition, and need not to carry out too much experiment.

Invention has been described in conjunction with specific embodiments, is appreciated that to do more the modification to it.The application is intended to cover substantially according to principle of the present invention and comprises and depart from the disclosed content of the present invention and as known in the field under the present invention or conventional practice and can be fit to the essential feature mentioned above that following claims scope is set forth, any change, use or adaptation that the present invention is done.

All documents that this description is quoted, comprise journal of writings or summary, open or the undocumented U.S. or foreign patent application, the U.S. that has announced or foreign patent or any other document, all intactly introduce this description as a reference, all data, form, accompanying drawing and the text that provide in the citing document are provided.And its full content of document of quoting in this description institute citing document also intactly draws and is reference.

In a word, with reference to known method step, conventional method step, known or conventional method, be not admit that any aspect of the present invention, any description or embodiment are disclosed in association area, instruction or prompting.

The foregoing description of specific embodiments will disclose general aspects of the present invention fully, other people can use this area knowledge (content that comprises this description institute citing document) and need not more experiment and just easily change and/or make these specific embodiments be applicable to various application, and can not deviate from total concept of the present invention.Therefore, these are suitable for and change means and drops on being equal in meaning and the scope based on the instruction of this description and the embodiment that discloses guided.The wording and the term that it should be understood that this description are the purposes unrestricted in order to describe, so that the term of this description or wording are according to instruction of the present invention and guide the knowledge in conjunction with those skilled in the art to be made an explanation by those skilled in the art.

Embodiment 1: the disease model of animal and long-term pharmacology

On different OA and damaged cartilage disease model, tested the remarkable interior therapeutic effect of FGF-18 (170AA) use intraarticular (i.a.) administration.Treating total effective dose at the last susceptible of proof of variety classes (rat, Canis familiaris L.) is 3-40 μ g/ animal/week (i.a.).The result who obtains from above-mentioned animal disease model (OA and cartilage defect) is summarized as follows:

The pharmaceutical research summary info of table 1:FGF-18 (170AA)

The pharmaceutical research summary info (continuing) of table 2:FGF-18 (170AA)

^aBe discussed in detail below

Embodiment 1.1: rat meniscus tear OA model

Method:

Male Lewis rat (5-10/group) underwent operative causes right tear of meniscus of knee joint.The operation back began to give 0.3,1,3 or 10 μ g dosage (with the saline preparation) through the i.a. approach in 19-21 days, measured the pharmacodynamics effect relevant with dosage regimen.These accumulated doses are as (i) single doses, (ii) weekly (per injection be accumulated dose 1/3) totally 3 weeks, or (iii) weekly 3 times, inject for 3 weeks (per injection be accumulated dose 1/9).When treatment finishes or after treatment 3 weeks of back, collect the histopathological evaluation that right knee joint is made potential effect.Lesion assessment carries out at 3 different district's bands: each section divides 3 district's bands (the 1-outside, 2-middle part, 3-inboard), takes into account the area differentiation of striding medial tibial plateau.In the OA surgery models, the outside (z1) and middle part (z2) two 1/3rd gets involved the most serious usually, and there is slightly variation in inboard 1/3rd (z3).Treatment procedure table summary:

Experimental design

Animal (5-10/group), 2-3/cage, use isoflurane anesthesia, prepare right knee joint operative region.At the inboard skin that cuts of knee joint, the exposed through blunt dissection medial collateral ligament, cross-section then.Cutting off the whole thickness of medial meniscus tears fully with simulation.With 4-0 Vicryl suture with the closed skin of suture way under the epidermis and subcutaneous tissue.Operation back stopped administration through the intraarticular administration in 19-21 days behind the single dose, or continued 3 all intra-articular injections, once in a week or 3 times weekly.Then rat is put to death (1-16 group) or makes its recovery, after 3 week back execution (17-32 group).In postmortem preceding 4 hours, all rats were given 50mg/kgBRDU with the active cell of labelling propagation.Serum is made in blood sampling before being about to postmortem, and carries out the lavation of right knee joint synovial fluid.During postmortem, right (operation) knee joint is pruned away muscle and connective tissue, collect in 10% neutral buffered formalin.Removing patella is completely fixed the joint.Trachea, breastbone and ear sample are also placed formalin.

The result of operation back assessment during 6 weeks

Untreated matched group (assessment during 6 weeks of operation back)

The not treatment animal that meniscus tear arranged is put to death during 6 weeks after operation, finds the tibia cartilage degradation, and 2/3 place is the most serious in the tibial plateau outside, and inboard 1/3 is more not serious.Hyperosteogeny big (average 468 μ m).The thigh cartilage degradation is lighter, and is bigger than tibia degeneration parameter.Compare with the animal of being untreated with the animal that saline is handled weekly once, 3 district's tibia cartilage degradation total points are lower (16%) obviously, and depth ratio is lower (16%) obviously, and the zone of lateral of tibia survival cartilage is big (13%) obviously.Handle 3 times animal tibia 2 districts cartilage degradations scoring obviously lower (24%) weekly with saline, depth ratio is lower (13%) obviously, and the cartilage zone that can survive in middle and lateral tibial is (15% and 14%) greatly obviously.These differences in the matched group may be to anaesthetize repeatedly to influence the overall active result of animal, but can not get rid of repeatedly the probability of pump pickle by promoting that medium and chip are removed influences the damage order of severity.

Single dose treatment when 6 weeks (after operation assessment)

Give the animal of 10 μ g FGF-18 (170AA) because the increase of 3 district mucin disappearance and total cartilage degradation width obviously (16%) greatly.The scoring of animal hyperosteogeny and the measured value that give 10 μ g FGF-18 (170AA) obviously increase (being respectively 19% and 25%).The zone of the cartilage matrix of surviving in the animal medial tibial with 10 μ g treatment obviously increases by 27%.Immunostaining has disclosed in bromine deoxidation urine sweet (BRDU) fibre modification bone marrow under the cartilage injury and the immune activeness in medullary cell, fibroblast and the hyperosteogeny cartilage, and pattern similarity dyes in the joint of saline and FGF-18 (170AA) processing.These results show that single injection 10 μ g FGF-18 (170AA) have clear and definite anabolic effect, as what proved by the increase of hyperosteogeny volume and the increase of medial tibial cartilage zone.But these deficient changes are to improve the scoring of all cartilage degradations.It seems that medial tibial cartilage zone increases be the result who increases near cartilage thickness on the load-bearing surface of hyperosteogeny.

Weekly 3 weeks for the treatment of when 6 weeks (after operation assessment)

The animal that gives 3 μ g FGF-18 (170AA) treatment is once in a week compared the marking of cartilage degradation in medial tibial 2 districts with the matched group that gives saline once in a week obviously lower.The obvious reduction that gives the animal degeneration width of 10 μ g FGF-18 (170AA) treatment has statistical significance (37%), and the animal that gives 3 μ g or 1 μ g also has some inhibition (being respectively 28% and 15%).Hyperosteogeny marking and measurement are dose response, are respectively 10 μ g FGF-18 (170AA) treatments (32 and 53%) or 3 μ g FGF-18 (170AA) treatments (21 and 32%).Obviously increase (27%) with 10 μ g FGF-18 (170AA) treatment great-hearted cartilage matrix zone in medial tibial.The intraarticular that injects 10 or 3 μ g have significantly slight to tangible chronic active synovial fluid with fibrosis, the intraarticular synovial fluid that injects 1 μ g is few.Animal BRDU labelling in Fibrotic bone marrow, synovial fluid and cartilage formation and hyperosteogeny zone with 3 or 10 μ gFGF-18 (170AA) treatment increases than saline control group.These results have proved to treat with weekly mode has the dose response effect to significant cartilage degradation width.This parameter is that the cartilage zone of defect area and medial tibial exists some type that the preferably indication of vigor substrate is arranged, and proves anabolic reaction.BRDU labelling result shows after the drug withdrawal lasting breeder reaction.

3 weeks for the treatment of when 6 weeks (after operation assessment) on every Wendesdays time

On every Wendesdays time injection 1,3 or 10 μ g FGF-18 (170AA) treat, and compare with time pump pickle on every Wendesdays, and the marking of (animal for the treatment of with 10 μ g), 2 districts (1 or 3 μ g) and 3 district's total amounts (1 or 3 μ g) tibia cartilage degradation obviously reduces in 1 district.Obviously reduce tibia cartilage degradation overall width 24% with 10 μ g treatment, reduce tibia cartilage degradation overall width 24% and 21% respectively but not remarkable on the statistics with 1 or 3 μ g treatment, showing has some useful effect.Animal with 1 or 3 μ g FGF-18 (170AA) treatment also has tangible depth ratio to reduce (being respectively 17% and 18%) than saline control group.After 0.3,1,3 or 10 μ g FGF-18 (170AA) treatment, the marking of medial tibial hyperosteogeny increases (being respectively 13%, 7%, 13% and 15%), and all 4 dosage (0.3,1,3 or 10 μ g FGF-18 (170AA)) all obviously increase hyperosteogeny measured value (being respectively 12,18,60 and 62%).Obviously increase the femur cartilage degradation with 10 μ g treatment and give a mark about 114%.With 3 (60%) or 10 (88%) μ g FGF-18 (170AA) treatment, bone marking obviously increases.With 3 μ g FGF-18 (170AA) treatment, the joint total points except femur reduces (13%) significantly, but femur is added to and has then eliminated this variation in the total points of joint.With 1,3 or 10 μ g FGF-18 (170AA) treatment, obviously increasing has vigor cartilage zone (13%, 29% and 29%) in the medial tibial, obviously vigor cartilage zone (22% and 13%) is arranged in the increase lateral tibial with 3 or 10 μ g treatment.Inject the visible serious synovitis in joint of 3 or 10 μ g, and subchondral bone absorbs and increases, inject the similar variation of the visible slight or moderate of 1 μ g in the joint.With the animal of 1,3 or 10 μ g FGF-18 (170AA) treatment, in a lot of zones (comprising bone marrow, hyperosteogeny, meniscus, synovial membrane and chondrogenic zone territory) the BRDU labelling is arranged all.These results have proved that any therapeutic modality all has the most tangible anabolic effect, but absorb increase with serious synovial fluid inflammation and subchondral bone.

The result of operation back assessment during 9 weeks

Do not treat matched group when week (after operation 9 assessment)

The not treatment animal that medial meniscus is torn was put to death when 9 weeks, and visible tibia cartilage degradation is the most serious in the outside 2/3 of tibial plateau, and is more not serious inboard 1/3.The femur cartilage degradation is more not serious, and vigor is arranged.The not treatment animal of putting to death when scoring generally was higher than for 6 weeks.The 2 district's cartilage degradation marking (12%) of single dose saline control group tibia are lower than weekly saline control group (18%).The width of single dose saline control group moderate cartilage degradation is obviously big (92%) also.The animal that gives saline once in a week is serious, obviously, the summation of moderate and slight cartilage degradation obviously lower (15%), and saline control animal on every Wendesdays time is 22%.Because every group only has 5 rats, these differences are less relatively, and this is the result of individual progression of disease difference.

Single dose (9 when week assessments after operation comprise treatment three weeks of back)

Give single dose FGF-18 (170AA) and 9 weeks put to death after operation any animals all do not have remarkable treatment effect, in lateral of tibia anabolic reaction is arranged although inject the joint of 10 μ g on a small quantity.

Weekly three weeks for the treatment of (assessment during 9 weeks comprises treatment three weeks of back after operation)

1 district and the total cartilage degradation scoring of 3 Qu of observing medial tibial after the weekly treatment of 10 μ g FGF-18 (170AA) obviously reduce by 38 and 31%.0.3 comparing the scoring in 2 districts with weekly saline control, μ g FGF-18 (170AA) treatment obviously reduces.Animal cartilage degeneration width with 10 μ g FGF-18 (170AA) treatment obviously reduces (38%).Obviously reduce (22%) with 10 μ g FGF-18 (170AA) treatment depth ratio.Treating significantly equal extent ground with 3 or 10 μ g FGF-18 (170AA) increases hyperosteogeny scoring (25%), but the animal hyperosteogeny measured value that only gives 10 μ g obviously increases (53%).The animal femoral joint total points that gives 10 μ g FGF-18 (170AA) obviously reduces by 23%.No matter the animal that gives 10 μ gFGF-18 (170AA) still is that lateral tibial has the area of vigor cartilage all obviously to increase (being respectively 40% and 81%) in the inboard.Treat the width that obviously reduces serious and minimum collagenous degeneration with 10 μ g FGF-18 (170AA), and serious, obvious and the width of moderate degeneration combination and the width of serious and obvious combination.The intraarticular synovial fluid inflammation of injecting 3 or 10 μ g is minimum, does not have the synovial fluid inflammation during lower dosage.Synthetic reaction appears significantly in some or all joints of injecting any dosage.The BRDU labelling is mainly seen in bone marrow and the fibroblast of saline control group, increases (cartilage and hyperosteogeny) at the intraarticular witness marking that injects 10 μ g.These results show, the continuing of repair of cartilage/synthetic reaction surpasses during the treatment, although suitable hyperosteogeny measured value is arranged during 6 or 9 weeks in the joint of this dosage treatment.As measured with collagenous degeneration, lack visible dose response effect improving with tangible cartilage degradation parameter and serious substrate.

Three weeks for the treatment of (assessment during 9 weeks comprises treatment three weeks of back after operation) on every Wendesdays time

10 μ g FGF-18 (170AA) inject the comparison on every Wendesdays time of three times animal and saline control weekly, and 3 districts tibia degeneration total points is reduction by 38% obviously.Also there is statistical significance ground obviously to reduce cartilage degradation width (48%) with 10 μ g FGF-18 (170AA) treatment.Animal depth ratio with 10 μ g FGF-18 (170AA) treatment also obviously reduces (27%).Measured value with the animal hyperosteogeny of 3 or 10 μ g FGF-18 (170AA) treatment obviously increases (being respectively 57% and 103%).Obviously increase the zone (being respectively 34%, 37% and 71%) that medial tibial has the vigor cartilage with 1,3 or 10 μ gFGF-18 (170AA) treatment, and obviously increase the zone (being respectively 46% and 67%) that lateral tibial has the vigor cartilage with 0.3 or 10 μ g FGF-18 (170AA) treatment.Animal mild or moderate collagenous degeneration width with 3 or 10 μ g FGF-18 (170AA) treatment obviously increases, and also is like this altogether at the width of the animal mild or moderate that gives 10 μ g and minimum degeneration.There is slight synovitis in the intraarticular that injects 10 μ g (gradation), and the intraarticular that injects 1 or 3 μ g exists and to be minimal to slight synovitis.From 1 μ g tangible synthetic reaction is arranged upwards.The BRDU labelling is mainly seen in bone marrow and the fibroblast of saline control group, increases (cartilage and hyperosteogeny) at the intraarticular witness marking that injects 10 μ g.These results show that the lasting of repair of cartilage/synthetic reaction surpasses during the treatment, compares synovitis with 6 time-of-weeks point and disappears.Useful effect to the collagen disappearance is not obvious, although there have clear and definite tendency to show to be obviously less to the measured value in serious disappearance zone.

Embodiment 1.2: Canis familiaris L. meniscotomy OA model

Female beagle (3/group), previous month in the up one-sided mid portion meniscotomy of left knee at begin treatment, then with saline or 3,10 or 30 μ g FGF-18 (170AA) carry out once, time (1/3 dosage that separates) treatment in totally three weeks once in a week or on every Wendesdays, measure the useful effect to the OA that has set up.

After treating for three weeks, assess variation that left knee perusal (3/group) and microscopic examination (3/organize) meniscotomy causes and the sign of synthetic reaction.In whole research process, except a Canis familiaris L., all dog foods desires and activity are all normal.Owing to anaesthetize the aspiration pneumonitis that causes repeatedly for joint injection, (the 17th day) death before research finishes of a Canis familiaris L. (YLI-8) of the 12nd group.In all groups all Canis familiaris L.s that undergo surgery all to occur on medial tibial with kitchen range limitation cartilage degradation damage be that the typical degenerative of feature changes.The femur damage is accidental.All Canis familiaris L.s all have the joint capsule medial thickness.Similar meniscus injury (about 1/2 disappearance during postmortem, none reparation or appropriateness are repaired usually) is all arranged in all groups.Microscopic evaluation discloses, and the knee for the treatment of and excipient treatment does not have the cartilage hypertrophy usually, and the cloning of level 1 and 2 is arranged in 1 district.Observe definite synthetic reaction (damage field increases cloning, cellularity and proteoglycan dyeing) with the joint of 30 μ g (10 μ g are inferior on every Wendesdays) FGF-18 (170AA) treatment, these variations are the most easy to identify at the femur ankle.With observing less but still compellent synergistic effect on every Wendesdays time or on weekly some joints of 30 or 10 μ g than low dosage FGF-18 (170AA) treatment.These variations that FGF-18 (170AA) causes when existing (if) are usually on cartilage in 1/3-1/2 or the contiguous substrate than deep crack.

The result of this research proved with 10 μ g FGF-18 (170AA) i.a. treatment on every Wendesdays time (30 μ g total amount/week) definite synergistic effect is arranged in all knees, use than low dose treatment on every Wendesdays time or in some knees for the treatment of once in a week with 30 or 10 μ g synergistic effect less.These variations are included in last 1/3-1/2 or the vicinity of cartilage and synthesize increase than the cloning in the substrate of deep crack and proteoglycan.In some joint, observe the marginal zone and slightly breed variation, do not go up the excessive variation that occurs but be similar to rat.The anabolism that anabolism in the articular cartilage changes greater than all case marginal zones changes.Collagen damage measurement value is also pointed out some protective effect of substrate integrity.

Embodiment 1.3: Canis familiaris L. anterior cruciate ligament (ACL) model

For the seriously research of carrying out property osteoarthritis, Canis familiaris L. anterior cruciate ligament (ACL) model is used for the research of preclinical pharmacology, comprises the treatment end and follows up a case by regular visits to middle MRI assessment.This model provides efficacy data (histopathology and time dependent MRI and follow up a case by regular visits to) and by the assessment data of gait analysis to function.

Read cartilage injury's reduction (excipient, 3 μ g/ joints, 10 μ g/ joints and 30 μ g/ joints are respectively-13.3 ,-7.5 ,-9.3 and-8.8) that the provable and healthy baseline of MRI is compared at the treatment end with Non-Invasive.Improve also provable with the function that the gait analysis for the treatment of end on assay plate measured.

Gait analysis adopt the pressure/force measuring instrument be installed on platform (

System, TekscanInc, Boston, MA USA) measures.

The gait capture program

System comprises 4 passage flat faced displays, and each display has 2,288 sensing elements that are included in 432mm * 368mm sensitizing range, produces 1.4 sensels/cm ²Spatial resolution.This equipment is proofreaied and correct with predetermined weight before beginning one's study, and all Canis familiaris L.s are all used same correction in the whole research.

For the hind leg that brings out osteoarthritis, with 1.9-2.2m/ Trotting gait speed acquisition vertical force and the contact area peak value of second.Guarantee speed with timer.The gait acquisition window is 3 seconds, and sample rate is located at 44hz, and common property is given birth to 132 frameworks.Every Canis familiaris L. obtains 5 initial efficiency tests.The feature of on average representing this Canis familiaris L. of particular point in time then.The vertical force peak value meter is shown the percent (%BW) of body weight, and contact area is expressed as square centimeter (cm ²).

The vertical force peak value that brings out the osteoarthritis hind leg that obtains during about quick step is compared with value (baseline) before the operation, and the Os Canitis arthritis model produces abnormal gait (the 4th week).All groups are all observed and are reduced in 4 weeks of operation back and 8 all vertical force peak values and contact area, are negative change (table 3 and 4).But there is the reduction of less severity the Canis familiaris L. operation back of group IV than other group during 8 weeks.

The vertical force peak value that brings out the osteoarthritis hind leg that obtains during table 3 quick step (dynamic item)

A. numeric representation is meansigma methods ± standard error, and numerical value is the change on baseline.

B. numeric representation is the percent (%BW) of body weight.

The contact area of bringing out the osteoarthritis hind leg that obtains during table 4 quick step (dynamic item)

A. numeric representation is meansigma methods ± standard error, and numerical value is the change on baseline.

B. numeric representation is the square centimeter (cm of body weight ²).

External pharmacology test result proves, dissimilar cellular exposure, and are not bred such as leukocyte or the side effect of release of cytokines at the sp act (propagation and cartilage degradation/collagen are synthetic) of chondrocyte in FGF-18 (170AA) back.

Embodiment 1.4: use radio-labeled FGF18[3H]-research that FGF18 carries out

(concentration of gross activity is expressed as ng equivalent/g) and table 6 (being expressed as the percent of dosage) has reported that intraarticular gives the radioactive level in [3H]-FGF18 patella for table 5; All reported the radioactivity that records in the joint of being treated with complete joint and collection synovianalysis in these tables.Table 7 has been reported corresponding pharmacokinetic parameter in the knee joint (have or do not have).

Table 5

With 0.24mg/kg target dose level give male rat intraarticular single give [ ³H]-the AS902330 patella in total radioactivity concentration.The result is expressed as ng equivalent/g.

Table 6

With 0.24mg/kg target dose level give male rat intraarticular single give [ ³H]-the AS902330 patella in the recovery of gross activity.The result is expressed as the percent of dosage.

Table 7

With 0.24mg/kg target dose level give male rat intraarticular single give [ ³H]-the AS902330 patella in system's exposure parameter of gross activity.

	The 3rd group of asynovia	The 4th group of whole joint	Ratio asynovia/whole joint
				C max(ngeq/g)	11740	26037	0.45
The recurrence scope	4-48	4-48
				t1/2，z(hr)	16	20
AUC(0-t At last)(ngeq·h/gr)	242116	393063	0.62
				AUC(0-∞)(ngeq·h/gr)	275787	492380	0.56

Being treated in the joint gross activity level is two-phase and reduces that (t1/2 is 20 hours.Take into account the whole joint that does not have synovial fluid, obtain comparable numerical value (16 hours).In a word, the result shows increased radioactivity outside synovial fluid, enters the knee joint tissue.The radioautographic analysis gained result who is treated the joint shows that most of radioactivity are positioned at next-door neighbour's joint.

Table 8 and table 9 reported respectively intravenous injection and intra-articular injection [ ³H]-whole-body autoradiograph analysis behind the FGF-18.Total radioactivity concentration in tissue and the organ is expressed as ng equivalent/g (meansigma methods ± SD).

Table 8

With 0.24mg/kg target dose level give the male rat single intravenous injection [ ³H]-total radioactivity concentration behind the AS902330 (the 1st group).The result is expressed as ng equivalent/g (meansigma methods ± SD).

^*NQ: beyond count (being lower than the quantitative limit)

Table 9

With 0.24mg/kg target dose level give male rat intraarticular single give [ ³H]-total radioactivity concentration behind the AS902330 (the 2nd group).The result is expressed as ng equivalent/g (meansigma methods ± SD).

^*NQ: beyond count (being lower than the quantitative limit)

After the intravenous injection 4 hours and 24 hours, the gross activity in blood and the serum reached peak respectively.The measured half-life is about 55 hours.After the intra-articular injection, the gross activity meansigma methods slowly raises in serum and the blood, respectively at reaching maximum concentration after the administration in 24 and 48 hours.System exposes and is about 20% of finding value after the intravenous injection.

Gross activity in the serum is higher than the gross activity in the blood on an average, and prompting parent compound and/or its metabolite are lower to the affinity of hemocyte, and the radioactivity in the circulation mainly is distributed in serum.

The gross activity level is the two-phase reduction in the subject joint, and t1/2 is 20 hours.In a word, the result shows that increased radioactivity is outside the synovial fluid that enters the knee joint tissue.

In a word, based on the non-clinical pharmacology model of rat and Canis familiaris L., find that specially suitable dosage regimen is totally three weeks once in a week.Observed effective dose changes between 3-30 μ g/ joint.

Embodiment 2: the example of FGF-18 chemical compound of the present invention

The sequence of preferred FGF-18 chemical compound of the present invention provides in following sequence table.

Sequence table

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＜120〉treatment of cartilage disorder

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Phe Arg Ile His Val Glu Asn Gln Thr Arg Ala Arg Asp Asp Val Ser

35 40 45

Arg Lys Gln Leu Arg Leu Tyr Gln Leu Tyr Ser Arg Thr Ser Gly Lys

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His Ile Gln Val Leu Gly Arg Arg Ile Ser Ala Arg Gly Glu Asp Gly

65 70 75 80

Asp Lys Tyr Ala Gln Leu Leu Val Glu Thr Asp Thr Phe Gly Ser Gln

85 90 95

Val Arg Ile Lys Glv Lys Glu Thr Glu Phe Tyr Leu Cys Met Asn Arg

100 105 110

Lys Gly Lys Leu Val Gly Lys Pro Asp Gly Thr Ser Lys Glu Cys Val

115 120 125

Phe Ile Glu Lys Val Leu Glu Asn Asn Tyr Thr Ala Leu Met Ser Ala

130 135 140

Lys Tyr Ser Gly Trp Tyr Val Gly Phe Thr Lys Lys Gly Arg Pro Arg

145 150 155 160

Lys Gly Pro Lys Thr Arg Glu Asn Gln Gln Asp Val His Phe Met Lys

165 170 175

Arg Tyr Pro Lys Gly Gln Pro Glu Leu Gln Lys Pro Phe Lys Tyr Thr

180 185 190

Thr Val Thr Lys Arg Ser Arg Arg Ile Arg Pro Thr His Pro Ala

195 200 205

<210>3

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acgcgggctc gagatgatgt gagtcggaag cagctgcgct tgtaccagct ctatagcagg 180

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1 5 10 15

Leu Leu Cys Phe Gln Val Gln Val Leu Ala Ala Glu Glu Asn Val Asp

20 25 30

Phe Arg Ile His Val Glu Asn Gln Thr Arg Ala Arg Asp Asp Val Ser

35 40 45

Arg Lys Gln Leu Arg Leu Tyr Gln Leu Tyr Ser Arg Thr Ser Gly Lys

50 55 60

His Ile Gln Val Leu Gly Arg Arg Ile Ser Ala Arg Gly Glu Asp Gly

65 70 75 80

Asp Lys Tyr Ala Gln Leu Leu Val Glu Thr Asp Thr Phe Gly Ser Gln

85 90 95

Val Arg Ile Lys Gly Lys Glu Thr Glu Phe Tyr Leu Cys Met Asn Arg

100 105 110

Lys Gly Lys Leu Val Gly Lys Pro Asp Gly Thr Ser Lys Glu Cys Val

115 120 125

Phe Ile Glu Lys Val Leu Glu Asn Asn Tyr Thr Ala Leu Met Ser Ala

130 135 140

Lys Tyr Ser Gly Trp Tyr Val Gly Phe Thr Lys Lys Gly Arg Pro Arg

145 150 155 160

Lys Gly Pro Lys Thr Arg Glu Asn Gln Gln Asp Val His Phe Met Lys

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Arg Tyr Pro Lys Gly Gln Ala Glu Leu Gln Lys Pro Phe Lys Tyr Thr

180 185 190

Thr Val Thr Lys Arg Ser Arg Arg Ile Arg Pro Thr His Pro Gly

195 200 205

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<212>PRT

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<220>

<223>FGF18(170AA)

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1 5 10 15

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20 25 30

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35 40 45

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50 55 60

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65 70 75 80

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85 90 95

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100 105 110

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115 120 125

Lys Lys Gly Arg Pro Arg Lys Gly Pro Lys Thr Arg Glu Asn Gln Gln

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Claims (22)

1.FGF-18 the application of chemical compound on preparation treatment cartilage disorder patient's medicine, wherein each treatment cycle intraarticular gives described FGF-18 chemical compound continuous at least 3 weeks, described dosing interval 6,7 or 8 days, and wherein said FGF-18 chemical compound is made up of SEQ ID NO:5.

2. application as claimed in claim 1, wherein said dosing interval are 7 days.

3. application as claimed in claim 1, wherein each treatment cycle gives described FGF-18 chemical compound continuous 3 week or continuous 4 weeks.

4. application as claimed in claim 2, wherein each treatment cycle gives described FGF-18 chemical compound continuous 3 week or continuous 4 weeks.

5. as each described application among the claim 1-4, wherein treatment cycle 2,4,6 or 8 months after repeat.

6. as each described application among the claim 1-4, wherein comprise 1,2,3,4,5 or 6 treatment cycle every year.

7. as each described application among the claim 1-4, wherein, for each intra-articular administration, the dosage of described FGF-18 chemical compound is 1-100mcg.

8. application as claimed in claim 7, wherein, for each intra-articular administration, the dosage of described FGF-18 chemical compound is 3,5,10,15,20,25,30,35,40,45,50,55 or 60mcg.

9. as each described application among the claim 1-4, wherein, described cartilage disorder is osteoarthritis.

10. application as claimed in claim 9, wherein, described osteoarthritis is categorized as slightly to the moderate osteoarthritis, is categorized as the osteoarthritis of II phase or III phase, serious carrying out property osteoarthritis and/or be categorized as the osteoarthritis of IV phase according to OARSI according to the international osteoarthritis OARSI of association.

11. application as claimed in claim 9, wherein said osteoarthritis are gonarthritis or coxitis.

12. application as claimed in claim 10, wherein said osteoarthritis are gonarthritis or coxitis.

13. application as claimed in claim 7, wherein, for each intra-articular administration, the dosage of described FGF-18 chemical compound is 5-40mcg.

14. application as claimed in claim 7, wherein, for each intra-articular administration, the dosage of described FGF-18 chemical compound is 10-30mcg.

15. application as claimed in claim 11, wherein said osteoarthritis are the Secondary cases coxitiss.

16. application as claimed in claim 12, wherein said osteoarthritis are the Secondary cases coxitiss.

17., it is characterized in that described cartilage disorder is the cartilage injury due to the wound as the described application of claim 1-4.

18. application as claimed in claim 1 is characterized in that, described FGF-18 chemical compound gives weekly once, continues for 3 weeks.

19. application as claimed in claim 6 is characterized in that, described FGF-18 chemical compound gives weekly once, continues for 3 weeks.

20. application as claimed in claim 7 is characterized in that, described FGF-18 chemical compound gives weekly once, continues for 3 weeks.

21. application as claimed in claim 9 is characterized in that, described FGF-18 chemical compound gives weekly once, continues for 3 weeks.

22. application as claimed in claim 17 is characterized in that, described FGF-18 chemical compound gives weekly once, continues for 3 weeks.