WO1994000145A1 - Targeted delivery of growth factors for bone regeneration - Google Patents
Targeted delivery of growth factors for bone regeneration Download PDFInfo
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- WO1994000145A1 WO1994000145A1 PCT/US1993/006254 US9306254W WO9400145A1 WO 1994000145 A1 WO1994000145 A1 WO 1994000145A1 US 9306254 W US9306254 W US 9306254W WO 9400145 A1 WO9400145 A1 WO 9400145A1
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- IOEWSPLILMTYPU-UHFFFAOYSA-N CC1=CC=CC(CNC)C1 Chemical compound CC1=CC=CC(CNC)C1 IOEWSPLILMTYPU-UHFFFAOYSA-N 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1858—Platelet-derived growth factor [PDGF]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/30—Insulin-like growth factors (Somatomedins), e.g. IGF-1, IGF-2
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/547—Chelates, e.g. Gd-DOTA or Zinc-amino acid chelates; Chelate-forming compounds, e.g. DOTA or ethylenediamine being covalently linked or complexed to the pharmacologically- or therapeutically-active agent
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/28—Phosphorus compounds with one or more P—C bonds
- C07F9/38—Phosphonic acids RP(=O)(OH)2; Thiophosphonic acids, i.e. RP(=X)(XH)2 (X = S, Se)
- C07F9/3804—Phosphonic acids RP(=O)(OH)2; Thiophosphonic acids, i.e. RP(=X)(XH)2 (X = S, Se) not used, see subgroups
- C07F9/3808—Acyclic saturated acids which can have further substituents on alkyl
- C07F9/3817—Acids containing the structure (RX)2P(=X)-alk-N...P (X = O, S, Se)
Definitions
- This invention relates to specific target delivery systems, namely, delivery of growth promoting factors to bone for its regeneration by a chelant composition system Compositions containing these systems and a process for making them are also part of this invention
- GF polypeptide growth factors
- IGF's insulin-like growth factors
- TGF transforming growth factors
- TGF- ⁇ 's beta's (TGF- ⁇ 's) and alpha (TGF- ⁇ )
- EGF epidermal growth factor
- FGF's fibroblast growth factor including acidic fibroblast growth factor (aFGF) and basic
- PDGF's are polypeptides of about 28-35 kilodaltons (kD) They are found in numerous cell types in the body PDGF derived from human platelets contains two polypeptide sequences, PDGF-A and PDGF-B polypeptides [See H N Antoniades and M Hunkapiller Sc/ence 220, 963-965 (1983)] PDGF-A is encoded by a gene localized in chromosome 7 [C Betsholtz et al , Nature 32Q_, 695-699 (1986)], and PDGF-B is encoded by the s/s oncogene [R Doolittle et al , Science 221, 275- 277 (1983), Waterfield et al , Nature 304, 35-39 (1983)] localized in chromosome 22 [R Dalla-Favera, Science 2J8, 686-688 (1982)]
- human PDGF occurs in three forms, a disulfide-lmked heterodimer of PDGF-A and PDGF-B, or two different homodimers (homodimer of PDGF-A and homodimer of PDGF-B)
- the role of PDGF in bone formation is not clear Some studies have indicated that it promotes bone resorption [Tashjian et al , Endocrinology 1 1 1 , 1 18-124 (1982), Canalis et al J Cell Physiol 140, 530-537 (1989)] Other studies have shown that PDGF stimulates the proliferation of osteoblasts in vitro and, when given via repeated subpe ⁇ osteal injections in newborn rats, new bone formation in vivo [Piche and Graves, Bone VO, 131-138 (1989), Joyce et al , in Clinical and Experimental Approaches to Dermal and Epidermal
- FGFs growth factors which have been studied for their effect on bone growth.
- aFGF and bFGF and their respective mRNA's have been detected at the site of bone fractures [Joyce et al., (1991 ) ibid].
- Both aFGF and bFGF have been isolated from bone matrix [Hauschka et al., (1986) ibid].
- bFGF and IGF-I have been used in combination to promote the healing of skin wounds [Lynch et al , J. Clm. Invest. (1989) ibid].
- bFGF did not significantly alter 3H-thym ⁇ dine incorporation in bone 0 fracture calluses [Joyce et al., (1991 ) ibid].
- bFGF has been reported to enhance mitogenesis in fetal calvarial bone cultures but did not simulate differentiated function of osteoblasts directly [Canalis et al., ;. Clm. Invest. 81, 1572 (1988)].
- aFGF has the same reported biological effects on bone as bFGF but generally requires higher concentrations [Canalis, J. Clin. Invest. 79, 52-58 (1987)].
- aFGF and bFGF tend to decrease matrix synthesis in the fetal rat calvarial model 5 [Canalis et al., (1989) ibid].
- Cultured bovine bone cells synthesize both bFGF and aFGF and store them in their extracellular matrix [Globus et al., Endocrinology 124, 1539 (1989)].
- bFGF has been reported to enhance the capacity of bone marrow cells to form bone-like nodules in vitro [Noff etaL. f.F. ⁇ .S. Letters 250. 619-21 (1989)].
- Both aFGF and bFGF increased DNA synthesis in cells cultured from parietal bones while bFGF was a more potent stimulator of alpha 1 Type 1 o procollagen mRNA [McCarthy et al., Endocrinology 125, 21 18-26 (1989)]. They are both mitogenic and chemotactic for cells derived from the pe ⁇ odontal ligament and bind to pretreated dentin slabs [Terranova et al., / Periodontol. 60, 293-301 (1989); Terra nova et al., J. Periodontol. 58, 247-257 (1987); Terranova, In The Biological Mechanisms of Tooth Extraction and Root Resorption, Davidovitch Z.
- TGF- ⁇ family of proteins also appear to have potential as modulators of bone growth.
- TGF- ⁇ is produced by osteoblasts and stimulates proliferation and collagen synthesis by these cells [Robey et al., J. Cell Biol. 105, 457-463 (1987); Rosen et al., Exper. Cell Res. 165, 127-138 (1986); Hock et al., Cal. Tissue Int. 32, 385 (abstract) (1988)].
- In vivo injections of TGF- ⁇ stimulate chondrogenesis and osteogenesis [Joyce et al., J. Cell Biol. 1 10, 2195-2207 0 (1990); Noda et al., Cndocr/no/oqy 124, 2991-2996 (1989)1
- Bone inductive factors such as bone morphogenetic proteins, osteogenin and osteoinductive protein 1 , can also stimulate bone formation They are often strucurally similiar to TGF- ⁇ and are characterized by their ability to induce ectopic cartilage and bone formation when implanted subcutaneously or intramuscularly in mammals (US Patents 4,877,864; 5 4,619,989; 4,455,256; 4,596,574; and 4,563,489; Wozney et al , S ⁇ ence 242, 1528-1534 ( 1988)] Targeted Delivery
- Current methods for delivery of proteins, such as, polymers, bone grafts and liposomes are unsatisfactory because they allow only localized delivery or systemic distribution without targeting.
- aminophosphonic and aminocarboxylic acid complexes having a metal complexed to an aminophosphonic or aminocarboxylic acid ligand, are known to deliver agents to bone [US Patents 4,508,704, 4,515,767, 4,560,548, 4,606,907, 4,897,254, 4,898,724, 5,059,412, 5,064,633, and 5,066,478].
- Most preferred would be the ability to deliver the desired growth factor(s) preferentially to the site of injured or depleted bone and allow for the activation of the bone growth factor(s) on an "as needed" basis as determined by the natural bone homeostasis mechanisms in the body.
- the present invention has the ability to met the above objectives.
- the present invention provides compounds for stimulating and enhancing bone growth by administering bone growth promoting factors which have been modified by being associated with a polyaminomethylenephosphonic acid ligand in a way that allows their preferential localization to skeletal tissue.
- the compounds for this delivery system are represented by the formula
- GF is a growth promoting factor or combinations thereof
- CL is an acid cleavable linker which is covalently bonded to GF; z is O, 1 or 2; q is from 1 to the sum of the amino groups present on the native GF;
- L is a linking moiety
- AP is a polyaminomethylenephosphonic acid ligand.
- Formulations for administering the compounds of Formula I to mammals, and methods forthe use of the compounds of Formula I for targeted delivery to bone, and processes for preparing the compounds of Formula I are also contemplated by this invention.
- compositions or formulations containing compounds of Formula I When the compositions or formulations containing compounds of Formula I are used, sites of injured or depleted bone are treated, and bone regenerated.
- Natural and recombinant tissue growth factors are commercially available from R&D Systems (Minneapolis, MN), Collaborative Research, Inc (Bedford, MA), Genzyme, Inc (Cambridge, MA), ICN Biomedicals, INC , (Cleveland, OH), Peprotech, Inc (Rocky Hill, NJ) and UBI (Lake Placid, NY) Bone inductive proteins can be purified from bone [Celeste et al , Proc Natl Acad Sci (USA) 87, 9843-9847 (1990), Wang et al , Proc Natl Acad So (USA) 85, 9484-9488 (1988), US Patents 4,455,256 and 4,619,989]
- GF growth promoting factors
- GF is chosen from PDGF's, IGF's, FGF's, TGF's or cartilage/bone inductive factors (BMP's)
- PDGF preferably in combination with IGF-I
- IGF-I has been shown to increase new bone formation when applied either alone or directly to diseased bone
- PDGF contains 30 free ammo groups per molecule which can potentially be modified to increase PDGF's affinity for bone PDGF is available from R&D Systems and Genzyme, Inc
- IGF-I has been described in US Patents 4,861 ,757 and 5,019,559
- IGF-I has been shown to increase new bone formation when applied, either alone or preferably in combination with PDGF, directly to diseased bone
- IGF-I contains 4 free ammo groups per molecule which can potentially be modified to increase PDGF's affinity for bone IGF-I is available from R&D Systems and Genzyme, Inc IGF-I is described by R E Humbel, Eur J Biochem
- IGF-II has been shown to increase new bone formation when applied directly to diseased bone IGF-II contains 2 free ammo groups per molecule which can potentially be modified to increase PDGF's affinity for bone IGF-II is available from R&D Systems and Genzyme, Inc IGF-II is described by R E Humbel, fur 7 Biochem 190, 445-462 (1990) bFGF has been shown to increase new bone formation when applied directly to diseased bone bFGF contains 15 free ammo groups per molecule which can potentially be modified to increase PDGF's affinity for bone bFGF is available from R&D Systems and Genzyme, Inc bFGF is described by Gospodarowicz et al .
- aFGF is a low molecular weight homodimer polypeptide which has been shown to increase new bone formation when applied directly to diseased bone aFGF contains 14 free ammo groups per molecule which can potentially be modified to increase PDGF's affinity for bone aFGF is available from R&D Systems and Genzyme, Inc aFGF is described by Gospodarowicz et al , Endocrmol Rev 8, 95-1 14 (1987)
- TGF- ⁇ is a low molecular weight (about 25 kDa, am o acids) homodimer polypeptide which has been shown to increase new bone formation when applied directly to diseased bone TGF- ⁇ ⁇ contains 18 free am o groups per molecule which can potentially be modified to increase PDGF's affinity for bone TGF- ⁇ ] is available from R&D Systems and Genzyme, Inc TGF- ⁇ , is described by Sporn et al , J Cell Biol 105, 1039-1045 (1987) BMP's have been shown to increase new bone formation when applied within mesenchymal tissues. The BMP's have been reviewed by Celeste et al. [Proc. Natl. Acad. Sci.
- the AP ligands may be straight or branched-chain moieties, cyclic moieties, polymers (including dense star polymers, their dendrimers and dendrons), or aryl moieties, which ligands contain at least two, preferably three or more, nitrogen atoms.
- the ligands are polyamino ⁇ methylenephosphonic acid ligands of one of the formula
- each R 1 independently is hydrogen, C.-C 4 alkyl, phenyl, hydroxy C ⁇ -C 4 alkyl, - CH 2 COOH, -CH-PO.H. or an L moiety; with the proviso that only one of R 1 may be an L moiety and one L moiety must be present and with the proviso that at least one-half of the total R 1 's are -CH-PO.H,; each R 2 and R 3 independently is hydrogen, C.-C, alkyl or L moiety; with the proviso that only one L moiety is present in Formula II; n is independently 2, 3 or 4; n' is independently 2, 3 or 4; and m is 0 to 10; or
- R 1 , R 2 , R 3 , n and m are defined as before; or
- R 1 is defined as before.
- the linking moiety (L in Formula I) is represented by the formula
- G is hydrogen, NH 2 or
- R 4 is an electrophilic group capable of being attached to protein
- R 5 and R 6 are independently hydrogen or -COOH; with the proviso that when G is hydrogen, then one of R 5 or R 6 is COOH; R 7 is hydrogen, hydroxy or C ⁇ C ⁇ alkoxy; and y is O, 1, 2, 3 or 4; with the proviso that when y is 1 , 2, 3 or 4, then only one of R 5 or R 6 may be COOH.
- straight or branched-chain moieties refers to an alkyl group having from 1 to about 100 carbon atoms which may be either a straight-chain moiety such as, for example, ethyl, propyl, n-butyl, n-dodecane and the like, or a branched-chain moiety such as, for example, isopropyl, tert-butyl, 2,5,7-trimethyldodecyl and the like. Both the straight and branched-chain moieties must contain at least 2 nitrogen atoms, preferably from 3 to 50 nitrogen atoms, and more preferably from 3 to 25. Some examples of these moieties include PDTMP
- PDTMP (N-propylcarboxyl)ethylenediamine-N,N',N'-trimethylenephosphonic acid
- APEDTMP [N-(4-aminophenyl)ethyl]ethylenediamine-N,N',N'-trimethylene- phosphonic acid
- CEDTMP 1 -(carboxyl)ethylenediamine-N,N,N',N'-tetramethylenephosphonic 0 acid
- ABEDTMP [1-(4-aminobenzyl)]ethylenediamine-N,N,N',N'-tetramethylene- phosphonic acid
- APIPTMP N-(4-aminophenyl)-N,N-bis-[propyl(iminodimethylenephosphonic acid)]; 5
- cyclic moieties refers to aliphatic, saturated ring systems having at
- aryl moieties refers to an aromatic ring system which may have one or more additional cyclic or aromatic rings or substitution by straight or branched-chain moieties
- the total number of atoms in the backbone of the aryl ring is from 3 to 30, preferably from 6 to
- the aryl moiety contains at least 2 nitrogen atoms, preferably from 3 to 10, and more preferrably from 3 to 8 Some examples of these moieties include pyrazolyl, 3-methylpyrazolyl, 5-methylpyrazolyl, imidazolyl, 4-methyl ⁇ m ⁇ dazolyl, 5- methylimidazolyl, 1 ,4-d ⁇ methyi ⁇ m ⁇ dazolyl .
- STARBU RSTTM dendrimers have polyammoimme groups where the surface has the ammo groups converted to aminomethyienephosphonic acid groups ana with at least one 4-am ⁇ nopheny!
- STARBURSTTM dendrimers are prepared as described in European Appln 0 271 180, published June 15, 1988, the disclosure of which is hereby incorporated by reference
- the "polymers” also include arborols (G R Newkome, V Org Chem. 50, 2004-2006 (1985), and either linear or branched polymer containing amines (V P Torchilin et al , Byull Eksp Biol Med. 102, 63-65 (1986)
- polyaminomethylenephosphonic acid [having a bifunctional chelantmg agent (BFCA) on the linking moiety (L of Formula I)] and the protein of the GF is reversible or cleavable under certain physiological conditions.
- BFCA bifunctional chelantmg agent
- Such linkages are taught in the art and include linkages contamg thiourea, thioether, peptide, ester, and difulfide groups [C F Meares et al., Int'l. J. Cancer, Supp 2, 99-102 (1988) and references contained therein, US Patent
- linkages between molecules and proteins which contain an amidine linkage are prepared by reacting a molecule containing an imidoester with the amine groups of the protein [O. R. Zaborsky, Immobilized Enzymes in Food and Microbial Processes, p187-202, A C. Olson and C C. Cooney, eds , Plenum, New York (1974)]
- group present to permit coupling to a protein or "electrophilic
- 35 group(s) capaole of being attached to prote ⁇ n(s) refers to an electrophilic group (R 4 of the L term of Formula I) that can bind to an ammo acid of a protein, e g the GF
- electrophilic groups include, but are not limited to, am o, maleimido, diazo, isothiocyanato, vinylpy ⁇ do, bromoacetamido, carboxyl, and N-hydroxy- succinimido active ester.
- polyaminomethylenephosphonic acid moieties are represented by a wide variety of possible groups, such as the cyclic moieties, straight or branched-chain moieties, aryl moieties, polymers including dense star poiymers, as defined above, which have at least one portion of the moiety containing between two nitrogen atoms a methylene (-CH 2 -) ⁇ group where n is 2, 3 or 4 (polyaminomethylene group). More than one such polyaminomethylene group may exist in the moiety.
- the moiety also contains at least two methylenephosphonic acid group (-CH 2 -P0 3 H 2 ) covalently attached to the polyamino- methylene group via a nitrogen.
- the polyaminomethylenephosphonic acid moieties are preferably represented by Formula II, III or IV but not limited thereto.
- the polyaminomethyl ⁇ enephosphonic acid moieties preferably have a group present to permit coupling to a protein or electrophilic group(s) capable of being attached to protein(s) as described herein.
- the polyaminomethylenephosphonic acid moieties (represented by L-AP in Formula I) may be linked to the protein (GF in Formula I) by a cleavable linker (represented by CL in Formula I), preferably an acid cleavable linker, as described herein.
- growth factor and "tissue growth promoting factor” mean any molecule which stimulates the proliferation, differentation, metabolism or migration of mammalian cells.
- the factors can be derived from natural sources or made by recombinant DMA technology or chemical synthesis. Preferably the factors are purified.
- purified refers to factors which, priorto mixing with the other growth factors, are 90% or greater, by weight, of the specified protein (i.e., is substantially free of other proteins, lipids, and carbohydrates with which it is naturally associated).
- a purified protein preparation will generally yield a single major band on a polyacrylamide gel for each subunit.
- the purified factor used in the compositions of the invention is pure as judged by amino-terminal amino acid sequence analysis.
- ligands of this invention may be in the form of their pharmaceutically acceptable salts.
- ligand as used herein is understood to include these salts.
- pharmaceutically acceptable salt means a cation acceptable for pharmaceutical use. These are cations that are not substantially toxic at the dosage administered to achieve the desired effect.
- these salts include those of alkali metals, such as sodium and potassium; alkaline earth metals, such as calcium and magnesium; ammonium; light metals of Group IIIA including aluminum; and organic primary, secondary and tertiary amines, such as trialkylamines, including triethylamine, procaine, dibenzylamine, N,N'-dibenzylethylenediamine, dihydroabiethylamine, N-(C 1 -C 4 )alkyipiperidine, and any other suitable amine.
- Sodium and potassium salts are preferred.
- pharmaceutically *' acceptable means suitable for administration to warmblooded animals, e.g.
- the pharmaceutically acceptable salts of the compounds of the present invention are prepared by conventional ion exchange processes or by treating the ligand or compound of Formula I with an appropriate base.
- ETMP ethylenediaminetetramethylenephosphonic acid
- EDA ethylenediamine
- EDTA ethylenediaminetetramet ylenecarboxyl c acid
- Open chain polyaminomethylenephosphonic acid BFCAs containing the linkage through an aminocarboxylic acid can be prepared from the corresponding polyamme intermediate BFCAs as described in European Appln 0 279 307, published August 24, 1988, the disclosure of which is hereby incorporated by reference.
- US Patents 4,994,560, 5,006,643 and 5,064,956 also describe how to make various open chain and cyclic amine containing BFCAs which can be phosphonomethylated to yield the corresponding polyaminomethylenephos ⁇ phonic acid BFCAs. Cyclic polyamine BFCAs are taught in EP Appln. 0 353450, published February 7,
- the cyclic BFCAs based on the 1 , 4,7,10-tetraazacyclododecyltetra- methylenephosphonic acid and the open chain BFCAs based on diethylenetriaminopenta- methylenephosphonic acid are a preferred group of polyaminomethylenephosphonic acid BFCAs ligands.
- the attachment group from the polyaminomethylenephosphonic acid BFCAs to the protein may be substituted from the cyclic polyaminomethylenephosphonic acid itself and are exemplified by J. P. L. Cox in 7. Chem. Soc, Chem. Commun. 797 (1989), and M. K. Moi et al, in J. Amer. Chem. Soc. 1_H), 6266-6267 (1988). Similar compounds are shown in WO 89/1 1475, published November 30, 1989.
- Cyclic amine compounds based on the 1 ,4,8, 1 1-tetraazacyclotetradecane have been described by M. K. Moi et al, in Inorg. Chem. 26, 3458-3463 (1987). These can be converted to the tetraaminomethylenephosphonic acid counterpart compounds by processes mentioned above. Similar compounds which may be used in the present invention are described in US Patent 4,678,667. Additional examples of cyclic polyamines are described by T. J. McMurry et al. in Bioconjugate Chem. 3(2), 108-1 17 (1992). These intermediate bifunctional cyclic amines can be phosphonomethylated to give the corresponding polyaminomethylene ⁇ phosphonic acid BFCA (the L-AP portion of Formula I).
- Bicyclopolyazamacrocylocarboxylic acid BFCA intermediates are disclosed in Kiefer et al.'s (assigned by unrecorded assignment to The Dow Chemical Company) copending US Patent Application Serial No. 805,270, filed December 10, 1991 , the disclosure of which is hereby incorporated by reference, can be phosphonomethylated to give the polyaminomethyl ⁇ enephosphonic acid BFCAs.
- Bicydopolyazamacrocylophosphonic acid BFCAs are disclosed in Kiefer et al.'s (assigned by unrecorded assignment to The Dow Chemical Company) copending US Patent Application Serial No. 805,551 , filed December 10, 1991 , the disclosure of which is hereby incorporated by reference, teaches the process to make various bicy opolyazamacrocylophos- phonic acid BFCAs having polyaminomethylenephosphonic acid BFCAs.
- BFCAs Bifunctional Chelating Agents having their Amino Groups Converted to Electrophilic Groups
- the BFCAs of the ligands of the present invention e.g. L of Formula I
- the process to convert the amino group to electrophilic groups capable of being attached to a protein is well known in the art. Some references that provide suitable process are: C. F. Meares et al., Ace. Chem. Res. V7, 202-209 (1984) and references given therein; D. Parker, Chem. Soc. Rev. 19, 271-291 (1990) and references given therein; C. F.
- Another method for linking two nucleophiles together is to convert one, such as an amine into a Michael acceptor, by reaction with acryloyl chloride orthe chemical equivalent. This converts the amino group to the acrylamide group which can then react with a different nucleophilic amine (Chem. Abst. 83:80295b).
- bifunctional crosslinking agents The molecules for attaching two nucleophilic moieties together are termed bifunctional crosslinking agents. If the two reactive ends of the molecule are the same, they are termed “homobifunctional” crosslinking agents. If the two reactive ends of the molecule are different, they are termed “heterobifunctional” crosslinking agents.
- Proteins and small molecules containing electrophilic groups such as amines can be modified by reaction with commercially available Traut's reagent [I. Wower, Nuc. Acid Res. 9, 4285-4291 (1981 )] to convert ammo groups to sulphydryl groups which can then react selectively with maleimide groups
- linkage between the polyaminomethyl ⁇ enephosphonic acid BFCAs and the GF be reversible or cleavable under certain physiological conditions.
- Such linkages are taught in the art and examples have been given above.
- Particularly advantageous are the linkers that are acid cleavable.
- Some examples of acid cleavable linkers are disclosed in US Patents 4,542,225 and 4,618,492 and the references mentioned therein. These cleavable linkers possess a cyclic anhydride at one end of the linker which reacts with amine groups, and a maleimido group at the other end of the linker which reacts with sulphydryl groups.
- the addition of metal ions does not significantly inhibit the affinity of polyaminomethylenephosphonic acids for calcific surfaces
- the addition of calcium or samarium ions does not interfere with the affinity of 1 , 4,7,10-tetraazacyclodo- decanetetramethylenephosphonic acid (DOTMP) for the calcific surface of hydroxyapitite
- DOTMP 4,7,10-tetraazacyclodo- decanetetramethylenephosphonic acid
- DOTMP has been shown to go to calcific sites in vivo when complexed with ions of samarium, holmium, gadolinium or yttrium in US Patent 4,976,950.
- ethylenediaminetetramethylenephosphonic acid (EDTMP) when complexed with calcium or other metal ions has also been shown to go to calcific sites in vivo (European Appln. 0462 787, published December 27, 1991)
- Polymers having polyaminomethylenephosphonic acids including dense star polymers such as described in European Appln. 0 272 180, published June 15, 1988, are prepared by reacting the amine with formaldehyde and dialkyl phosphite in an aqueous solution which forms the peralkyl phosphonate ester. This ester is then hydrolyzed to the aminomethylenephosphonic acid on the dense star polymer.
- the dense star is in the PAMAM form having amine groups on its surface. These amine groups are then reacted with phosphonomethylating agents to form the polyaminomethylenephosphonic acid, which is then conjugated to the GF.
- the advantages of the dense star as the bone seeking moiety are that it can be water soluble, have a controlled size, and specific groups and quantity of groups available for conjugation to GF.
- the polymers of the L-AP of Formula I also include arborols (G. R. Newkome, J. Org. Chem. 50, 2004-2006 (1985) which are capable of being attached to proteins, e.g. GF.
- the arborols are monocascade spheres which possess a three-deminsional microenviroment having the outer surface covered with polar functional groups.
- These arborols can be converted into polyamines by methods known in the art for converting esters and alcohols into amines (e.g. Survey of Organic Synthesis 1 , 41 1-417 (1970), pub. John Wiley and references given therein).
- the arborol polyamines can be converted into polyaminomethylenephosphonic acids by the methods described above.
- the arborols can contain a group capable of attaching to proteins.
- aborols containing polyaminomethylenephosphonic acid groups and a group capable of attaching to a protein by either a stable covalent linkage or cleavable linkage can be prepared.
- Selective attachment of the polyaminomethylenephosphonic acid BFCAs to noncritical amine groups on a GF may be accomplished by first mixing the GF with a soluble or solid support immobilized form of the receptor from cells which will form a GF-receptor aggregate. This aggregate can then be reacted with an excess of the reactive form of the poly- aminomethylenephosphonic acid BFCA. Since the GF is bound to the receptor, only amine groups not involved in the GF receptor interaction will be modified. The GF modified in this fashion is then able to interact with its receptor v vo to yield biological activity. The aggregate between the modified GF and the receptor can then be broken down into the now selectively modified GF and the reusable receptor, which after separation will yield fully active modified GF that still recognizes the receptor.
- the compounds of Formula I can be prepared in several different ways.
- the AP portion can be reacted with L to form the L-AP portion; the L-AP portion then attached to the CL; with the CL-L-AP group then attached to the GF.
- GF is reacted with CL to form the GF-CL portion; which GF-CL is then reacted with L-AP to form compounds of Formula I.
- the degree of modification of the GF is described by the q term of Formula I, which is arrived at experimentially by adjusting the concentrations, time, temperature, pH, and stoichiometry of the reactions between GF, CL and the L-AP molecules.
- the formulations of the present invention are in the solid or liquid form. These formulations may be supplied as a single substance for direct use or as two or more substances (e.g. in kit form) such that the two components are mixed at the appropriate time prior to use. Whether premixed or as a kit, the formulations may require a pharmaceutical ly-acceptable carrier or adjuvant.
- the compounds of this invention may also be administered parenterally, that is, subcutaneously, intravenously, intramuscularly, or interperitoneally, as injectable dosages of the compound in a physiologically acceptable diluent with a pharmaceutical carrier which can be a sterile liquid or mixture of liquids such as water, saline, aqueous dextrose and related sugar solutions, an alcohol such as ethanol, isopropanol, or hexadecyl alcohol, glycols such as propylene glycol or polyethylene glycol, glycerol ketals such as 2, 2-di methyl- 1 ,3-dioxolane-4- methanol, ethers such as poly(ethyleneglycol) 400, an oil, a fatty acid, a fatty acid ester or glyceride, or an acetylated fatty acid glyceride with or without the addition of a pharmaceutically acceptable surfactant such as a soap or a detergent, suspending agent such as pec
- Suitable fatty acids include oleic acid, stearic acid, and isostearic acid.
- Suitable fatty acid esters are, for example, ethyl oleate and isopropyl myristate.
- Suitable soaps include fatty alkali metal, ammonium, and triethanolamine salts and suitable detergents include cationic detergents, for example, dimethyl dialkyl ammonium halides, alkyl pyridinium halides, and alkylamines acetates; anionic detergents, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates; nonionic detergents, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylenepoly-propylene copolymers; and amphoteric detergents, for example, alkyl-
- compositions of this invention will typically contain from about 0.001 % to about 10% by weight of compound of Formula I in solution. Preservatives and buffers may also be used advan-tageously. In order to minimize or eliminate irritation at the site of injection, such compositions may contain a non- ionic surfactant having a hydrophile-lipophile balance (H LB) of from about 12 to about 17.
- H LB hydrophile-lipophile balance
- the surfactant can be a single component having the above HLB or can be a mixture of two or more components having the desired HLB.
- Illustrative of surfactants used in parenteral formulations are the class of polyethylene sorbitan fatty acid esters, for example, sorbita ⁇ monooleate.
- compositions of Formula I include: orally, using known oral pharmaceutical formulations some of which may contain magnesium hydroxide in excessive amounts [P. J.Neuvonen et al., fur. J. Clin. Pharmacol. 35, 495-501 (1988)]; trans- dermal delivery using known procedures from B. Kari, Diabetes 35, 217 (1986), B. R. Meyer et al., C/ n. Pharmacol. Ther. 44, 607 (1988); implants using adsorption of the compound onto activated carbon particles which are then implanted or injected [A. Hagiware, Gan to Kagaku Ryoho V5, 1038-1042 (1988) or Chem. Abst.
- formulations may also be applied locally to the site of the injured or depleted bone by direct topical application to that site. The latter method may require surgery to expose the site of injured or depleted bone.
- Bone regeneration using the compounds of the present invention is more effective than that achieved in the absence of treatment (i.e. without applying exogenous agents) or by treatment with similar levels of unmodified growth promoting factors, i.e. only GF of Formula I. Because of the relatively high cost of GF production and the potential ability of GF to cause adverse toxic affects when delivered in high doses systemically, it is also desireable to use the ligand to ensure delivery to the desired site and reduce the overall dose of GF and its toxic effects to the mammal.
- the amount of (CL).-L-AP of Formula I which is needed to aid in bone delivery of the GF can be any effective amount.
- the amount of compound of Formula I to be administered in order to treat any of the diseases desired for such a delivery system can vary widely according to the particular dosage unit employed, the period of treatment, the age and sex of the patient treated, the nature and extent of the disorder treated, and other factors well-known to * those practicing the medical arts.
- the compounds of Formula I can be used in conjunction with other agents known to be useful in the treatment of bone diseases.
- the effective amount of compounds of Formula I to be administered according to the present invention will generally range from about 0.005 to 50 mg/kg of body weight of the patient and can be administered as frequently as one or more times per day.
- the compounds of Formula I can be administered in a pharmaceutically acceptable formulation as described above.
- the compounds of Formula I can have one or more different active compounds administered either simultaneously or sequentially; and such compounds may be administered with other known active agents for regenerating bone.
- the invention will be further clarified by a consideration of the foil owing examples, which are intended to be purely exemplary of the present invention. Definitions
- BMP bone morphogenetic proteins
- BSA bolvine serum albumin
- FCS fetal calf sera
- HSA human serum albumin
- PBS phosphate buffered saline
- SCN-BDTMP 4-isoth ⁇ ocyanatobenzyldiethylenetr ⁇ am ⁇ nopentamethyienephos- phonic acid
- Acetic acid was glacial acetic acid from Ald ⁇ ch Chemical Co. at greater than
- Both PDGF-BB and IGF-I were produced by standard recombmant DNA techniques and purified by conventional chromatography Both process are well known to those ski 11 in the art (PDGF- US Patents 4,061 , 757 and 5,045,633, IGF-I Y Sato et al , ./ Biochem 101 , 1247- 1252 (1987) and Wong et al , Gene 68, 193-203 (1988)
- the cells are preincubated with PDGF-BB, prior to the addition of test samples and standards.
- the premcubation enables the cells to respond to IGF-I when the IGF-I is added in the presence of Epidermal Growth Factor (EGF).
- EGF Epidermal Growth Factor
- a unit is defined that concentration of GF which induces 50% of the maximum cellular response in the appropriate assay (ED50 value).
- Maximum response in the PDGF asnd IGF-I assays is defined as the cellular response to a 5% Fetal Calf Sera (FCS) standard.
- FCS Fetal Calf Sera
- the specific activity was calculated by dividing the number of units based on the mitogenic assay by the mass as determined by amino acid analysis.
- the potency for both PDGF-BB and IGF-I prior to protein modification was 1 to 3 ng/mL (see Figure 3 for PDGF-BB; Figure 4 for IGF-I).
- the specific activities were in the range of 3 to 10 x 10 5 units per mg of protein.
- Example C Radiolabelled 1 2 5
- the "cold" PDGF described in Example A was labelled by the [ 12 5
- 125 I-PDGF is also commercially available from NEN. NEN normally supplies the 125 l- PDGF in a sodium citrate buffered salt solution containing 1 % BSA as a carrier protein Prior to use this BSA must be removed from the commercially available ,25 I-PDGF to allow efficient modification of the PDGF In order to produce the carrier free form of 1 5
- the immobilized anti-PDGF antibody (supplied by 1MB) exhibits specific binding to the AB and BB PDGF isoforms, and no binding to the AA homodimer 42.6 ⁇ Ci 12 5
- the BSA was exhaustively flushed from the column with PBS and the 1 5
- Example D Radiolabelled 125
- the "cold " IGF described above was labelled with [ 125 l] using the lactoperoxidase or chloramme T method which lodmatestyrosine residues forming lodotyrosyl products
- 125 l- IGF-I is commercially available from NEN 1 2
- -IGF-I was supplied carrier free and freeze dried from 100 uL of 100mM sodium citrate, pH 4 5 Based on a specific activity of 208 ⁇ C ⁇ / ⁇ g, this lot of 125 I-IGF-I is estimated to have on average, one [ 125 l] attached for every 1 4 IGF-I protein chains
- Example E Preparation of two PDGF solutions
- a vial containing radioactive PDGF was placed in the well of a Capmtec dose calibrator which was adjusted for 1 $
- a vial containing 50 ⁇ g of PDGF was rinsed with 200 ⁇ L of 0 1 M acetic acid and added to a vial containing 100 ⁇ g of cold IGF and 1 5
- the cold PDGF was vial was rinsed with 7 more 200 ⁇ L portions of 0 1 M acetic acid, which rinses were combined to provide one sample containing radioactive and nonradioactive PDGF and nonradioactive IGF
- the combined sample vial was shaken to mix and two 750 ⁇ L aliquots of the solution were removed Each aliquot was placed on a separate membrane of a centncon 10, labelled 1 and 2 Appropriate volume and gamma readings were taken which verified the volume and radioactivity before and after dividing of the sample The two aliquot samples were spun in a centrifuge
- the ester 250 mg (0 3 mmol) was hydrolyzed by stirring with 3 mL of concentrated HCI at 100°C for 18 hours The aqueous solution was freeze-d ⁇ ed to give (80%), as a cream colored solid, 1-( ⁇ -carboxyl-2-methoxy-5-n ⁇ trobenzyl)-1 ,4,7, 10-tetraazacyclodo- decane-4,7, 10-tr ⁇ methylenephosphon ⁇ c ac ⁇ d (O N-BD03TMP) A 100 mg portion of 0 2 N-BD03TMP was dissolved in 20 mL of water. After purging the system with nitrogen, 120 mg of 10% Pd/C was added and the suspension placed under an atmosphere of hydrogen with constant vigerous stirring.
- HPLC anion exchange, eluting with O to 1 M ammonium acetate over 30 minute 0 gradent a 2 mL/min
- HPLC indicated a slight shift to a shorter retention time for the reduced product.
- the suspension was then filtered, and the filtrate lyophilized to give 259 mg of white glassy solid.
- HPLC indicated a purity of about 92% .
- the product was further purified by preparative anion exchange chromatrography as described above to give, as a white solid, 144 mg of the ammonium salt of 1 -(4-aminobenzyl)diethyienetriaminepentamethylenephosphonic acid, with 5 a purity by anion exchange HPLC of >95% .
- the product was further characterized by as follows. Decoupled P-31 NMR showed 3 singlets in the expected 221 ratio
- the recombmant IGF vial's contents were dissolved in 10mM of acetic acid and transferred into the glass serum vial containing lodmated IGF dissolved in 535 mg of 10mM acetic acid
- This combined solution of IGF containing tracer 125 I-IGF-I was dialysed (Spectra/Por7TM membrane from Spectrum Medical Industries) against three 1 L portions of
- Frozen radiolabelled 125 l-lGF as a 150 ⁇ L aliqout containing 300 ⁇ g of IGF-I and 23 6 ⁇ C ⁇ of carrier free 125 I-IGF in 0 1M acetic acid was allowed to thaw and then diluted to a final volume of about 1 5 mL using 0 1 M acetic acid A 700 ⁇ L aliqout of this solution was
- This compound may be used as a ligand AP in Formula I where the L is through the COOH group.
- Example K
- the flask was attached to a reflux condenser, equipped with a stir bar, and set on o a stirrer
- a 10 mL syringe was filled with 17 74 g (0 219 moles) of 37% formaldehyde solution and attached to a syringe pump pre-calibrated to deliver at a flow rate of 0 1 mL/min
- the reaction solution was brought up to reflux temperature, then with constant stirring, the formaldehyde was slowly added to the flask over a three hour period
- This compound may be used as a ligand AP in Formula I where the L is through the NH 2 group
- Example L Preparation of 1 -(Carboxy)ethylened ⁇ am ⁇ netetramethylenephosphon ⁇ c Ac ⁇ d (CEDTMP) The structure of the compound CEDTMP is shown below
- This compound may be used as a ligand AP in Formula I where the L is through the NH 2 group
- Example N Preparation of N 4 -(p-am ⁇ nophenyl)-norsperm ⁇ d ⁇ ne
- This compound may be used as a ligand AP in Formula I where the L is through the NH 2 group
- APIPMP N"(4-am ⁇ nophenyl)-d ⁇ propylenetr ⁇ am ⁇ ne-N',N',N'",N'"- tetramethylenephosphonic Acid
- N 4 -(p-am ⁇ nophenyl)norsperm ⁇ d ⁇ ne prepared in Example N (3 65 g, 16 4 ⁇ moles) was added to 25 mL of water and treated with a solution of 6 2 g (75 6 mMoles) phosphorous acid in 13 mL (163 mMoles) of concentrated HCL The resulting dark blue solution was heated to 100°C and treated dropwise with 5 9 g (73 mMoles) of 37 weight percent formaldehyde in water over a 2 75 hour period The solution was kept at reflux for an additional 15 hours, then cooled to room temperature and dripped into ethyl alcohol The resulting brown precipitate was vacuum filtered and dried under vacuum to give 3 84 g (40% yield) of the title compound (APIPMP) as a brown solid The product thus obtained gave a retention time of 5 50 minutes (minor peak) and 10 50 (major peak) when analyzed on the anion exchange HPLC system described in Example L
- This compound may be used as a ligand AP in Formula I where the L is through the NH 2 group
- Example P
- Diethylenet ⁇ ami ⁇ e (20 g, 0.045 mole) was dissolved in 200 mL of toluene and treated over five minutes with a solution of p-nitrophenethyl bromide (10 g, 0.89 mole) in 150 mL of toluene. Afterthree hours of stirring the supernatent was decanted from the gummy solid and extracted three times with 100 mL portions of water. The combined water layers were reduced under vacuum to a low volume and back extracted with 100 mL chloroform. The chloroform was then evaporated under vacuum to yield 9.32 g (83%) of N"-(p-n ⁇ trophenethyl)- diethylenetriamine.
- This compound may be used as a ligand AP in Formula I where the L is through the NH 2 group.
- Example Q
- the 4-isothiocyanatophthalic acid thus obtained was immediately heated to reflux in a mixture of trifluoroacetic anhydride and methylene chloride for 2 hours under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure. Recrystallization of the resultant solid from 30 mL of carbon tetrachloride afforded 4-isothiocyanatophthalic anhydride as brownish-purple crystals in a yield of 2.3778 g, (77 percent of theoretical).
- Example 1 A 400 ⁇ L portion of 34.7mM SCNBDTMP solution (1.389 x 10 "5 moles) (prepared in Example G) was added to aliquot 1 from Example E. This steoichometry gives a 794: 1 molar ratio of isothiocyanate group per amino group on the protein (Sample 1 ).
- CentriconTM no 2 of Example I (containing approximately 127 ⁇ g of IGF)
- the solution residing on the CentriconTM membrane was then mixed well and allowed to set for 13 hours at room temperature
- the unit was then spun to concentrate the protein of the solution
- the residual on the CentriconTM membrane was then removed by adding 855 ⁇ L of a 1 % HSA solution, swirrlmg and pipetting.
- IGF modified with BDTMP using ACL-3 A 210 ⁇ L aliquot of IGF solution ( 100 ⁇ g of IGF containing tracer amounts of 1 5
- a 100 ⁇ L aliquot of PDGF solution was put on the membrane of a CentriconTM membrane filtration device
- a 500 ⁇ L aliquot of 0 3M sodium bicarbonate was added to the CentriconTM unit and it was then centrifuged until 50 mg remained on the membrane
- the contents on the membrane (washed PDGF) were taken up in 500 ⁇ L more of 0 3M sodium bicarbonate
- the membrane retained material was then treated with the 36 7 mg of ACL- 3 dissolved in 300 ⁇ L of warm trifluoroethanol, allowed to stand for 15 minutes, and then centrifuged for 2 hours At the end of this time a 300 ⁇ L volume of ABH DP (357
- Spraque-Dawley rats weighing in the 175-230 g weight range were acclimated for 5 5 days prior to injection.
- the rats were injected via the tail vein with 50 ⁇ L of both samples from Example 1 (Samples 1 and Comparative A) (about 200,000 cpm).
- the rats were sacrificed by cervical dislocation, tissues taken, weighed and the amount of radioactivity in each tissue determined by counting for 5 min. in a scintillation counter equipped with a Nal crystal coupled to a multichannel analyzer. The counts in each tissue were o compared to the counts in 50 ⁇ L standards in order to determine the percentage of injected dose in each tissue.
- Spraque-Dawley rats weighing in the 175-230 g weight range were acclimated for 5 days prior to injection.
- the rats were injected via the tail vein with 50 ⁇ L of one of the four samples from Examples 2, 3, 4 or Comparative B. After 30 mins. the rats were sacrificed by cervical dislocation, tissues taken, weighed and the amount of radioactivity in each tissue
- the percent dose in bone was estimated by multiplying the percent dose in the femur by 25.
- the muscle and blood numbers were obtained assuming that muscle is 43% and blood is 6.5% ⁇ of the rat body weight. The results are given in Table II where each data point represents the average of five rats.
- Spraque-Dawley rats weighing in the 175-230 g weight range were acclimated for 5 days p ⁇ orto injection
- the rats were injected via the tail vein with 50 ⁇ L of the samples from Examples 5 or C
- the rats were sacrificed by cervical dislocation, tissues taken, weighed and the amount of radioactivity in each tissue determined by counting for 5 mm in a scintillation counter equipped with a Nal crystal coupled to a multichannel analyzer
- the counts in each tissue were compared to the counts in 50 ⁇ L standards in order to determine the percentage of injected dose in each tissue
- the counts in the tail were subtracted from the standards to give the percent injected dose adjusted for the amount found in the tail Background counts were obtained and subtracted from the tissue counts
- the percent dose in bone was estimated by multiplying the percent dose in the femur by 25
- the muscle and blood numbers were obtained assuming that muscle is 43% and blood is 6 5% of the rat body weight
- a unit is defined as that concentration of IGF-I which induces 50% of the maximum cellular response in that assay (ED 50 value)
- Maximum response is defined as cellular response to a 5% FCS standard
- Maximum response due to exogenous IGF-I is usually 3-5 times baseline P ⁇ orto dilution for the assay, the concentration of IGF-I was determined via an
- Example 5 sample was 107.2 ⁇ g/mL; and Example C was 1 10.8 ⁇ g/mL. The results are shown in Table V.
- Example V 25 l-IGF-ACL-3-BDTMP
- Doses (150 ⁇ L) of the material from Example 7 and Comparative Example E were drawn up into 500 ⁇ L syringes equipped with 28 gauge needles and injected into the tail veins of 5 rats. After 6 hours the rats were sacrificed and dissected to obtain the organ distribution of the radiolabelled material. Because of the low amount of radioactivity involved, the samples were counted for 10 minutes. The final number of counts (less background) found in the femurs of rats injected with the modified IGF material of Example 7 were 3.8 times the number of counts (less background) found in the femurs of rats injected with the control material of Comparative E as seen in the following Table VI.
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
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JP6502666A JPH07508979A (en) | 1992-06-30 | 1993-06-30 | Targeted transport of growth factors for bone regeneration |
EP93916894A EP0653941A1 (en) | 1992-06-30 | 1993-06-30 | Targeted delivery of growth factors for bone regeneration |
AU46600/93A AU4660093A (en) | 1992-06-30 | 1993-06-30 | Targeted delivery of growth factors for bone regeneration |
FI946156A FI946156A (en) | 1992-06-30 | 1994-12-29 | Targeted supply of growth factors that affect bone regeneration |
NO945093A NO945093L (en) | 1992-06-30 | 1994-12-30 | Targeted release of growth factors for bone regeneration |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
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US90698092A | 1992-06-30 | 1992-06-30 | |
US07/906,980 | 1992-06-30 | ||
US08/026,800 US5505931A (en) | 1993-03-04 | 1993-03-04 | Acid cleavable compounds, their preparation and use as bifunctional acid-labile crosslinking agents |
US08/026,800 | 1993-03-04 |
Publications (1)
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WO1994000145A1 true WO1994000145A1 (en) | 1994-01-06 |
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PCT/US1993/006254 WO1994000145A1 (en) | 1992-06-30 | 1993-06-30 | Targeted delivery of growth factors for bone regeneration |
Country Status (10)
Country | Link |
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EP (1) | EP0653941A1 (en) |
JP (1) | JPH07508979A (en) |
CN (1) | CN1092076A (en) |
AU (1) | AU4660093A (en) |
CA (1) | CA2139323A1 (en) |
FI (1) | FI946156A (en) |
HU (1) | HUT71220A (en) |
IL (1) | IL106159A0 (en) |
MX (1) | MX9303963A (en) |
WO (1) | WO1994000145A1 (en) |
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0711300A4 (en) * | 1993-05-06 | 1996-03-12 | Dow Chemical Co | 2-pyridylmethylenepolyazamacrocyclophosphonic acids, complexes and derivatives thereof, for use as contrast agents |
US5776193A (en) * | 1995-10-16 | 1998-07-07 | Orquest, Inc. | Bone grafting matrix |
GB2322129A (en) * | 1997-02-13 | 1998-08-19 | Ciba Sc Holding Ag | Polyazabicyclic compounds |
WO2001028567A2 (en) * | 1999-10-18 | 2001-04-26 | The Dow Chemical Company | Aminoalkylenephosphonates for treatment of bone disorders |
WO2001077102A1 (en) * | 2000-04-07 | 2001-10-18 | Bristol-Myers Squibb Pharma Company | Macrocyclic chelants for metallopharmaceuticals |
US6794371B1 (en) * | 1999-10-18 | 2004-09-21 | The Dow Chemical Company | Aminoalkylenephosphonates for treatment of bone disorders |
WO2004089356A2 (en) * | 2003-04-03 | 2004-10-21 | Semafore Pharmaceuticals Inc. | Targeted bone marrow protection agents |
US6902584B2 (en) | 1995-10-16 | 2005-06-07 | Depuy Spine, Inc. | Bone grafting matrix |
EP1932850A1 (en) * | 2006-12-11 | 2008-06-18 | Thermphos Trading GmbH | Phosphonate compounds |
WO2016123454A1 (en) | 2015-01-29 | 2016-08-04 | Board Of Trustees Of Miching State University | Cryptic polypeptides and uses thereof |
US10071182B2 (en) | 2014-10-14 | 2018-09-11 | Samuel E. Lynch | Methods for treating wounds |
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JPH02268190A (en) * | 1989-04-07 | 1990-11-01 | Fujisawa Pharmaceut Co Ltd | Conjugate from medical compound and diphosphonic acid derivative |
US5011913A (en) * | 1985-06-28 | 1991-04-30 | The Procter & Gamble Company | Diphosphonate-derivatized macromolecules |
EP0512844A1 (en) * | 1991-05-10 | 1992-11-11 | Celtrix Pharmaceuticals, Inc. | Targeted delivery of bone growth factors |
US5208219A (en) * | 1991-02-14 | 1993-05-04 | Celtrix Pharmaceuticals Inc. | Method for inducing bone growth |
-
1993
- 1993-06-28 IL IL106159A patent/IL106159A0/en unknown
- 1993-06-30 AU AU46600/93A patent/AU4660093A/en not_active Abandoned
- 1993-06-30 CN CN93109549A patent/CN1092076A/en active Pending
- 1993-06-30 JP JP6502666A patent/JPH07508979A/en active Pending
- 1993-06-30 HU HU9403840A patent/HUT71220A/en unknown
- 1993-06-30 MX MX9303963A patent/MX9303963A/en unknown
- 1993-06-30 EP EP93916894A patent/EP0653941A1/en not_active Withdrawn
- 1993-06-30 WO PCT/US1993/006254 patent/WO1994000145A1/en not_active Application Discontinuation
- 1993-06-30 CA CA002139323A patent/CA2139323A1/en not_active Abandoned
-
1994
- 1994-12-29 FI FI946156A patent/FI946156A/en unknown
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US5011913A (en) * | 1985-06-28 | 1991-04-30 | The Procter & Gamble Company | Diphosphonate-derivatized macromolecules |
JPH02268190A (en) * | 1989-04-07 | 1990-11-01 | Fujisawa Pharmaceut Co Ltd | Conjugate from medical compound and diphosphonic acid derivative |
US5208219A (en) * | 1991-02-14 | 1993-05-04 | Celtrix Pharmaceuticals Inc. | Method for inducing bone growth |
EP0512844A1 (en) * | 1991-05-10 | 1992-11-11 | Celtrix Pharmaceuticals, Inc. | Targeted delivery of bone growth factors |
Cited By (21)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0711300A1 (en) * | 1993-05-06 | 1996-05-15 | The Dow Chemical Company | 2-pyridylmethylenepolyazamacrocyclophosphonic acids, complexes and derivatives thereof, for use as contrast agents |
EP0711300A4 (en) * | 1993-05-06 | 1996-03-12 | Dow Chemical Co | 2-pyridylmethylenepolyazamacrocyclophosphonic acids, complexes and derivatives thereof, for use as contrast agents |
US6764517B2 (en) | 1995-10-16 | 2004-07-20 | Depuy Acromed, Inc. | Tissue repair matrix |
US5776193A (en) * | 1995-10-16 | 1998-07-07 | Orquest, Inc. | Bone grafting matrix |
US6187047B1 (en) | 1995-10-16 | 2001-02-13 | Orquest, Inc. | Bone grafting matrix |
US7842097B2 (en) | 1995-10-16 | 2010-11-30 | Depuy Spine, Inc. | Tissue repair matrix |
US6902584B2 (en) | 1995-10-16 | 2005-06-07 | Depuy Spine, Inc. | Bone grafting matrix |
GB2322129A (en) * | 1997-02-13 | 1998-08-19 | Ciba Sc Holding Ag | Polyazabicyclic compounds |
US6794371B1 (en) * | 1999-10-18 | 2004-09-21 | The Dow Chemical Company | Aminoalkylenephosphonates for treatment of bone disorders |
WO2001028567A3 (en) * | 1999-10-18 | 2001-11-29 | Dow Chemical Co | Aminoalkylenephosphonates for treatment of bone disorders |
WO2001028567A2 (en) * | 1999-10-18 | 2001-04-26 | The Dow Chemical Company | Aminoalkylenephosphonates for treatment of bone disorders |
US6565828B2 (en) | 2000-04-07 | 2003-05-20 | Bristol-Myers Squibb Company | Macrocyclic chelants for metallopharmaceuticals |
WO2001077102A1 (en) * | 2000-04-07 | 2001-10-18 | Bristol-Myers Squibb Pharma Company | Macrocyclic chelants for metallopharmaceuticals |
WO2004089356A2 (en) * | 2003-04-03 | 2004-10-21 | Semafore Pharmaceuticals Inc. | Targeted bone marrow protection agents |
WO2004089356A3 (en) * | 2003-04-03 | 2004-11-25 | Semafore Pharmaceuticals Inc | Targeted bone marrow protection agents |
EP1932850A1 (en) * | 2006-12-11 | 2008-06-18 | Thermphos Trading GmbH | Phosphonate compounds |
WO2008071692A3 (en) * | 2006-12-11 | 2009-01-15 | Thermphos Trading Gmbh | Phosphonate compounds |
RU2537946C2 (en) * | 2006-12-11 | 2015-01-10 | Италматч Кемикалс СпА | Phosphonate compounds |
US9296632B2 (en) | 2006-12-11 | 2016-03-29 | Italmatch Chemicals Spa | Phosphonate compounds |
US10071182B2 (en) | 2014-10-14 | 2018-09-11 | Samuel E. Lynch | Methods for treating wounds |
WO2016123454A1 (en) | 2015-01-29 | 2016-08-04 | Board Of Trustees Of Miching State University | Cryptic polypeptides and uses thereof |
Also Published As
Publication number | Publication date |
---|---|
HUT71220A (en) | 1995-11-28 |
JPH07508979A (en) | 1995-10-05 |
MX9303963A (en) | 1995-01-31 |
FI946156A (en) | 1995-02-27 |
CN1092076A (en) | 1994-09-14 |
IL106159A0 (en) | 1993-10-20 |
EP0653941A1 (en) | 1995-05-24 |
HU9403840D0 (en) | 1995-02-28 |
FI946156A0 (en) | 1994-12-29 |
CA2139323A1 (en) | 1994-01-06 |
AU4660093A (en) | 1994-01-24 |
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