Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/74—Synthetic polymeric materials
  • A61K31/765—Polymers containing oxygen
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/74—Synthetic polymeric materials
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/74—Synthetic polymeric materials
  • A61K31/785—Polymers containing nitrogen
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/74—Synthetic polymeric materials
  • A61K31/795—Polymers containing sulfur
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system

Definitions

• the present invention relates to methods of therapy involving the use of said pharmaceutical compositions containing polyaromatic compounds having properties which mimic pharmacological activities of glycosaminoglycans and effect the distribution in tissue of biologically active peptides and proteins normally bound to glycosaminoglycans.
• Glycosaminoglycans are linear polysaccharides formed by characteristic repeating disaccharide units usually composed of a uronic acid and a hexosamine.
• the term "acid mucopolysaccharides” was used originally to designate hexosamine-rich acid polysaccharides extracted from connective tissue.
• the term “glycosaminoglycans” has gained greater acceptance and is now used in place of mucopolysaccharides.
• the hexosamine can be glycocyamine or galactosamine
• the uronic acid can be glucuronic or iduronic acid.
• glycosaminoglycans apart from hyaluronic acid, and all of the sulphated glycosaminoglycans are covalently linked to protein forming different classes of proteoglycans.
• glycosaminoglycans serve also as a support which binds various bioactive peptides. This association is based on a non-coyalent interaction since the bound protein can be readily released upon the addition of free glycosaminoglycans.
• bound proteins include enzymes such as lipoprotein lipase (LPL) or growth-regulating peptides such as fibroblast growth factor (FGF) .
• LPL lipoprotein lipase
• FGF fibroblast growth factor
• Another example of GAG-protein interaction is that of the enzyme heparinase which participates in cell-invasion processes.
• glycosaminoglycan heparin
• Heparin is known also to be involved in the release of lipoprotein lipase, the inhibition of heparinase and the release of fibroblast growth factor.
• the most common application of heparin is as an anticoagulant where heparin interacts with proteins which play a key role in hemostasis.
• Glycosaminoglycans such as heparin are a major constituent participating in the composition of various biological structures such as basement membranes, connective tissues, cartilage and cell-surface glycocalyx. Connective tissues are responsible for providing and maintaining form in the body. Functioning in a mechanical role, they provide a matrix that serves to connect and bind the cells and organs and ultimately give support to the body.
• connective tissue Unlike the other tissue types (epithelium, muscle and nerve) formed mainly by cells, the major constituent of connective -tissue is its extracellular matrix, composed of protein fibers, an amorphous ground substance, and tissue fluid, the latter consisting primarily of bound water of solvation. Embedded within the extracellular matrix are the connective tissue cells.
• connective tissue In terms of structural composition, connective tissue can be subdivided into three classes of components: cells, fibers and ground substance. The wide variety of connective tissue types in the body represents modulations in the degree of expression of these three components.
• the amorphous intercellular ground substance fills the space between cells and fibers of the connective tissue; it is viscous and acts as a lubricant and also as a barrier to the penetration of the tissues by foreign particles.
• Glycosaminoglycans and structural glycoproteins are the two principal classes of components comprising the ground substance.
• the present invention is based on the discovery of a class of compounds exhibiting properties which mimic the action of glycosaminoglycans and which are capable of modulating biological systems containing complexes between bioactive peptides and/or proteins and glycosaminoglycans by competing with the binding interactions of glycosaminoglycans.
• the present invention relates to a pharmaceutical composition
• a pharmaceutical composition comprising, in admixture with a pharmaceutically acceptable carrier, a therapeutically effective amount of a polyaromatic ring-containing polymeric compound, substantially free of monomer, and having properties which mimic the pharmacological activity of glycosaminoglycans and which are capable of competing with the binding thereof to bioactive peptides and/or proteins.
• the present invention relates more specifically to a pharmaceutical composition including as an active ingredient a therapeutically effective amount of a polyaromatic ring-containing polymeric compound having a molecular weight between about 2,000 and about 20,000 Daltons wherein each monomeric unit of said polymer includes from 1 to about 10 aromatic rings, which may be substituted by electronegative substituents and/or negatively charged residues.
• This invention relates also to therapeutic methods comprising the administration of an effective amount of the above-mentioned pharmaceutical composition to human or other animal patients in need of cardiovascular therapy such as anticoagulant and/or antithrombotic therapy and/or bone metabolic therapy and/or therapy for the treatment of neuronal disorders.
• cardiovascular therapy such as anticoagulant and/or antithrombotic therapy and/or bone metabolic therapy and/or therapy for the treatment of neuronal disorders.
• tissue distribution is used herein to mean either the compartmentalization of a molecular constituent within a given tissue or the distribution pattern between different tissues.
• tissue redistribution* means a change in the compartmentalization within a given tissue or a change in the distribution pattern between tissues.
• a peptide may be released from a basement membrane which can result subsequently in its consumption by surrounding cells or its transfer into the bloodstream, or a soluble compound could be bound to a protein in a competitive binding situation thereby preventing the association of said compound with certain tissue-fixed residues.
• Alkyl means a saturated aliphatic hydrocarbon which may be either straight or branched-chained containing from about 1 to about 12 carbon atoms.
• “Lower alkyl” means an alkyl group as above, having 1 to about 4 carbon atoms.
• Substituted phenyl means a phenyl group substituted with one or more substituents which may be alkyl, alkoxy, amino, nitro, carboxy, carboalkoxy, cyano, alkyl amino, halo, hydroxy, hydroxyalkyl, mercaptyl, alkyl mercaptyl, carboalkyl or carbamoyl.
• substituents may be alkyl, alkoxy, amino, nitro, carboxy, carboalkoxy, cyano, alkyl amino, halo, hydroxy, hydroxyalkyl, mercaptyl, alkyl mercaptyl, carboalkyl or carbamoyl.
• the polymeric compounds included in the compositions of this invention may be useful in the form of the free base, in the form of salts, esters and as hydrates. All forms are within the scope of the invention. Acid addition salts may be formed and are simply a more convenient form for use? and in practice, use of the. salt form inherently amounts to use of the base form.
• the acids which can be used to prepare the acid addition salts include preferably those which produce, when combined with the free base, pharmaceutically acceptable salts, that is, salts whose anions are non-toxic to the animal organism in phar ⁇ maceutical doses of the salts, so that the beneficial pharmacological properties inherent in the free base are not vitiated by side effects ascribable to the anions.
• Pharmaceutically acceptable salts of the compounds useful in the practice of this invention are those derived from the following acids: mineral acids such as hydrochloric acid, sulfuric acid, phosphoric acid and sulfamic acid; and organic acids such as acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, quinic acid, and the like.
• mineral acids such as hydrochloric acid, sulfuric acid, phosphoric acid and sulfamic acid
• organic acids such as acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic
• the corresponding acid addition salts comprise the following: hydrochloride, ⁇ ulfate, phosphate, sulfamate, acetate, citrate, lactate, tartarate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclohexylsul- famate and quinate, respectively.
• the acid addition salts of the compounds contained in the compositions of the present invention are prepared either by dissolving the free base in aqueous or aqueous-alcohol solution or other suitable solvents containing the appropriate acid and isolating the salt by evaporating the solution, or by reacting the free base and acid in an organic solvent, in which case the salt separates directly or can be obtained by concentration of the solution.
• the more preferred number of aromatic rings in each monomeric unit is about three to about six.
• Polymers of many known synthetic dyes comprising monomers having the polyaromatic characteristics identified above are within the scope of the present invention and are useful in pharmaceutical compositions. Such dyes are known to be useful as color additives, diagnostic aids administered by i.v. injection, antiseptic agents and/or to treat infectious disease. Nevertheless, none of these dyes has previously been described as useful as or used as a therapeutic agent as a replacement for glycosaminoglycans.
• Exemplary synthetic dyes including polymeric materials in accordance with this invention include Aluminon, Halogenated Alu inon, Sulfonated Aluminon, Sulfonated-Halogenated Aluminon, Anazolene Sodium, Eosine I Bluish, Eosine Yellowish, Erythrosine, Evan's Blue, Fast Green FCF, Fucshin(e) Acid, Iodophthalein Sodium, Rose Bengal, Sulfobromophthalein Sodium, Suramin Sodium, Trypan Blue, Trypan Red, Rosaniline Chloride, Crystal Violet, Methyl Blue, Methyl Green, Coomassie Blue, Basic Fuchsin, Malachite Green, Brilliant Green, Aniline Blue, Brilliant Cresyl Blue, Safranin 0, Ethyl Violet, Pararosaniline Acetate and Methyl Violet.
• substituent x, y, z
• a, b and c are independently 0 or 1 and a+b+c > 2 and m is about 5 to about 20.
• the preferred molecular weight of the polymeric compounds is about 2,000 and about 20,000, and the most preferred molecular weight is about 2,000 to about 4,000 Daltons, as measured by gel permeation chromatography.
• compositions containing the pharmacologically active polymeric compounds are believed to function according to one or more of the following mechanisms.
• FGF fibroblast growth factor
• Preferred embodiments of the present invention are described in the following non-limiting examples which include a description of pharmacological test procedures believed to correlate to therapeutic activity in humans and other animals.
• Comparative Compound A Commercially available aurintricarboxylic acid (ATA) from Aldrich Chemical Co., Wisconsin.
• This material is prepared using commercially available aurintricarboxylic acid which is fractionated on Biogel P-6 by elution with 50% aqueous ethanol with 50mM Tris at pH 7.5 and then 50% aqueous ethanol with 50mM glycine at pH 11.
• the fraction containing polymeric material having a molecular weight of about 2000 to about 4000 Daltons is separated.
• Comparative Compound C - ATA not containing polymers Following the procedure described in Organic Synthesis. Coll. Vol. I, page 54, the following reaction was performed to produce what is believed to comprise non- polymerized ATA. To 17 ml of concentrated sulfuric acid at 0-5°C and mechanically stirred was added portionwise sodium nitrite (2.5 g , 36.2 mmol) . The mixture was stirred until a solution was obtained. Salicylic acid (5.0 g , 36.5 mmol) was added portionwise over 10-15 minutes and the reaction warmed to room temperature, stirred until a homogeneous mixture was obtained and cooled to 0-5*C.
• Comparative Compound D - Polymer believed to contain ATA monomeric units is prepared by using a large excess of formalin in the above procedure along with the following reagents.
• the polymeric components of the following dyes significantly prolong the activated partial thromboplastin time when compared to a standard buffer solution.
• LPL participate in the process of lipid transfer from the bloodstream to the tissues, LPL is bound to the external surface of cells via its association with cell-membrane glycosaminoglycans.
• Cultured heart cells is a powerful system in the study of the enzyme turn-over and the exploration of ways to manipulate its level since these cells are active in the biosynthesis of LPL, in addition to the surface binding of the enzyme.
• F x heart cell cultures are prepared from newborn rate hearts as previously described (Chajek, T. et al., Biochem. Biophys. Acta 521, 456-474 (1978)). They consist mainly of non-beating mesenchymal cells and are used 8-12 days after their isolation.
• Enzyme activity is determined using aliquots of medium and of homogenates of cells which are released from a petri dish with a rubber policeman in 1 ml of 0.025 M NH 3 /NH 4 CI buffer (pH 8.1) containing 1 unit/ml of heparin.
• the assay system consists of 0.1 ml of medium or 0.1 ml of cell homogenate (50-70 g protein) and 0.1 ml of substrate containing labeled triolein, prepared according to the method of Nilsson-Ehle and Schotz, (J. Lipid Res. 12, 536-541 (1976)). Incubations are carried out at 37*C for 45 mins.
• the reaction is stopped by the addition of methanol/chloroform/heptane (1.4:1.25:l.v/v) and the extraction of fatty acids is performed according to the method of Belfrage and Vaughan (J. Lipid Res. 2SL, 341- 344 (1969), as modified by Nilsson-Ehle and Schotz. Enzyme activity is calculated according to the formula of Nilsson-Ehle and Schotz.
• compositions within the scope of this invention are useful in the treatment of cardiovascular diseases such as arteriosclerosis.
• the active ingredients in the composition of the present invention are effective inhibitors of heparinase activity.
• the inhibitory activity is measured by the following procedure. Inhibition of Heparinase Activity
• Heparinase is an endoglucuronidase capable of degrading heparin sulfate (HS) at specific intrachain sites.
• HS heparin sulfate
• ECM subendothelial extracellular matrix
• compositions of the present invention to inhibit lymphoma-cell derived heparinase is tested in the assay system described by Vlodavsky et al. (Cancer Research. 43:2704-2711 (1983)).
• 35 S labeled ECM is incubated for 24 hours with ESb mouse lymphoma heparinase in the presence of 10 ugr ml of the indicated compounds.
• Degradation of the sulfated glycosaminoglycan is followed by gel filtration of the supernatants.
• Heparinase activity is expressed as the total amount of labeled low-molecular-weight fragments released from the EMC substrate.
• Anticoagulant properties are evaluated in accordance with the following method.
• the anticoagulant effect of commercially available Aluminon is demonstrated by measuring the partial thromboplastin time (PTT) in blood samples from a normal donor. 0.25 ml of the indicated concentration of the drug (xlO, diluted in saline) is added to 2.25 ml of fresh human blood, collected in the presence of citrate. PTT is measured following the addition of Ca " " " .
• aluminon is an effective anticoagulant.
• LPL were elevated by about an order of magnitude.
• compositions of the present invention are capable of being administered orally to produce an anticoagulant effect.
• heparin can only be administered by injection.
• the active ingredients of the compositions within the scope of this invention are useful inhibitors of the proliferation of vascular smooth muscle cells.
• the proliferation inhibition of the active ingredients of the present compositions may be evaluated according to the following procedure. Inhibition of Vascular-Smooth Muscle Cell Proliferation
• Bovine aortal smooth-muscle cells are grown in tissue culture plates in the presence of glucose-rich medium (H-21) .
• the cultures (around 35xl0 3 cells/well in 24 wells' plates) are initially supplemented with 10% fetal calf serum (FCS) for 2-3 days.
• FCS fetal calf serum
• the cell nutrition is then replaced by a serum-deprived medium (H-21 plus 0.2% FCS) in order to test the effect of defined growth factors on the proliferation of smooth- muscle cells and their possible inhibition by the compounds with heparin-like activity.
• the cells transferred to the serum-deprived medium are grown in the presence of thrombin (10 "6 M) or FGF (fibroblast growth factor, partially purified from bovine brain, 250 ngr/ l) .
• thrombin 10 "6 M
• FGF fibroblast growth factor, partially purified from bovine brain, 250 ngr/ l
• Various concentrations of Aluminon were added to the thrombin or FGF-treated cells and to non-treated control cultures. 3 H thymidine incorporated into DNA was measured. The results are presented in Table V below.
• compositions of the present invention are useful in the treatment of cardiovascular diseases, including arteriosclerosis, bone metabolism and neuronal disorders.
• compositions of this invention can be normally administered orally or parenterally, in the treatment of cardiovascular disorders, bone metabolic disorders and neuronal disorders in humans or other mammals.
• compositions of this invention may be formulated for administration in any convenient way, and the invention includes within its scope pharmaceutical compositions containing at least one polymeric compound as described hereinabove adapted for use in human or veterinary medicine.
• Such compositions may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers or excipient ⁇ .
• Suitable carriers include diluents or fillers, sterile aqueous media and various non-toxic organic solvents.
• compositions may be formulated in the form of tablets, capsules, lozenges, troches, hard candies, .powders, aqueous suspensions, or solutions, injectable solutions, elixirs, syrups and the like and may contain one or more agents selected from the group including sweetening agents, flavoring agents, coloring agents and preserving agents, in order to provide a pharmaceutically acceptable preparation.
• the particular carrier and the ratio of therapeutically effective compound to carrier are determined by the solubility and chemical properties of the compounds, the particular mode of administration and standard pharmaceutical practice.
• excipients such as lactose, sodium citrate, calcium carbonate and dicalcium phosphate and various disin- tegratants such as starch, alginic acid and certain complex silicates, together with lubricating agents such as magnesium stearate, sodium lauryl; sodium lauryl sulphate and talc, can be used in producing tablets.
• lactose and high molecular weight polyethylene glycols are among the preferred pharmaceutically acceptable carriers.
• the carrier can be emulsifying or suspending agents. Diluents such as ethanol, propylene glycol, glycerin and chloroform and their combinations can be employed as well as other materials.
• solutions or suspensions of the polymeric compounds in sesame or peanut oil or aqueous propylene glycol solutions, as well as sterile aqueous solutions can be employed.
• the aqueous solutions using pure distilled water are also useful for intravenous injection purposes, provided that their pH is properly adjusted, suitably buffered, made isotonic with sufficient saline or glucose and sterilized by heating or by microfiltration.
• the dosage regimen in carrying out the methods of this invention is that which insures maximum therapeutic response until improvement is obtained and thereafter the minimum effective level which gives relief.
• the dosages are those that are therapeutically effective in increasing the clotting time of blood, decreasing the chances of thrombosis, reducing the buildup of arterial plaque or in the treatment of bone etobolic or neuronal disorders such as Alzheimer's disease.
• the oral dose may be between about 1 mg/kg and about 500 mg/kg (preferably in the range of 1 to 10 mg/kg), and the i.v. dose about 0.1 mg/kg to about 10 mg/kg (preferably in the range of about 0.1 to about 3 mg/kg), bearing in mind, of course, that in selecting the appropriate dosage in any specific case, consideration must be given to the patient's weight, general health, age and other factors which may influence response to the drug.
• the drug may be administered as frequently as is necessary to achieve and sustain the desired therapeutic response. Some patients may respond quickly to a relatively large or small dose and require little or no maintenance dosage. On the other hand, other patients may require sustained dosing from about 1 to about 4 times a day depending on the physiological needs of the particular patient. Usually the drug may be administered orally 1 to 4 times per day. It is anticipated that many patients will require no more than about one to about two doses daily.
• the present invention would be useful as an injectable dosage form which may be administered in an emergency to a patient suffering from stroke or heart attack. Such treatment may be followed by intravenous infusion of the active polymeric compound and the amount of compound infused into such a patient should be effective to achieve and maintain the desired therapeutic response.

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• 1990
  • 1990-08-13 IL IL9535390A patent/IL95353A/en not_active IP Right Cessation
  • 1990-08-14 AU AU63375/90A patent/AU649963B2/en not_active Ceased
  • 1990-08-14 WO PCT/US1990/004580 patent/WO1991003226A2/en not_active Application Discontinuation
  • 1990-08-14 JP JP2512506A patent/JPH05503506A/ja active Pending
  • 1990-08-14 CA CA002064575A patent/CA2064575A1/en not_active Abandoned
  • 1990-08-14 EP EP19900913407 patent/EP0489102A4/en not_active Withdrawn

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