JPH10510831A - Complex of dermatan sulfate and drug to improve drug - Google Patents

Abstract

  (57) [Summary] Disclosed is a drug carrier composition containing a drug complexed with dermatan sulfate. The drug is preferably an anti-tumor agent, and can be taxol, a peptide tumor agent, or vincristine. The most preferred antineoplastic is doxorubicin. Dermatan sulfate is an essentially purified dermatan sulfate with a sulfur content of up to 9% (w / w) and selective sulfation of oligosaccharides. The composition allows for efficient vascular access and is administered in a manner that induces the following effects in vivo: 1) rapid, partial or total endothelial coating of the drug (diagnostic) carrier; ) If the endothelial pocket overlying the carrier has not yet penetrated into the vascular compartment, sequester the carrier quickly (2 minutes) and protect the captured drug from blood clearance; 3) vascular endothelium Or accelerate the transport of carriers into the tissue compartment (gap) across and / or through the subendothelial structure; and 4) increase the efficiency of the drug traveling through the endothelium or the barrier of the upper or subendothelial layer. By improving, a lower total drug dose is required to achieve the desired effect compared to the dose required for a standard drug. Similar tissue uptake is described for transepithelial migration to the lung, bladder, and intestine.

Description

DETAILED DESCRIPTION OF THE INVENTION             Complex of dermatan sulfate and drug to improve drug                               Background of the Invention   Until recently, the localization of intravascular drugs in body tissue has been Have relied on chemical partitioning of body organs into tissue compartments. This results in an injection dose that actually reaches the intended target of only 0.01% to 0.001. It was only%. About 20 years ago, drugs were captured in liposomes and microspheres (entrap) was done. This modifies the initial biodistribution and sets them apart from the following details: Phagocytic cells in reticulocellular organs were redirected: liver, spleen and bone marrow.   In 1978, the present inventors and co-workers (Widder et al., Proc. Am, Assn. Cancer Re. s., p. 19, p. 17 (1978)) are microspheres (which are non-reticulum cell targeting devices). It is injected intravenously into tissue compartments of government (eg, lung, and brain) and is magnetically localized. Developed a means to co-entrap drug and magnetite together . Magnetic capture is normal tissue, with sufficient strength (0.5 Tesla to 0.8 Tesla) and gradient (0 Tesla). .1 Tesla / mm) to the tumor tissue placed in close proximity to the extracorporeal magnet via the vascular endothelium. Achieved by selectively towing offspring. This technology is both efficient and The desired target tissue is placed at an injection dose between 25% and 50%, but it is also Was a very complex approach and had the following major disadvantages: 1) Restricted use to specialized medical center; 2) Target Permanent placement of magnetite in tissue; 3) inhomogeneity of magnetic field capture )) Resulting in local overdosing of the drug; and 4) extremely restricted Applies to a limited number of therapeutic agents. However, projects that study magnetic targeting Delay of toxic drugs from captured carriers (microspheres) (Controlled) release protects normal cells in the local tissue environment from drug toxicity. And still provided an effective treatment of tumor cells and microorganisms.   When monoclonal antibodies can be used for animal and clinical research in general, -Drug conjugates limit the biodistribution of toxic drugs and reduce them to disease foci ( Tumors and infections), which have been placed through the microvascular barrier in the target tissue It was desired to be placed. Unfortunately, most monoclonal antibodies are (And is still available) from a monoclonal antibody Is made immunologically foreign to the human recipient. For therapeutically relevant replacement ratios Drug conjugation makes monoclonal antibody derivatives even more foreign and To reduce their binding specificity. For this reason, antibody-drug conjugates It is substantially purified by the liver, as in Importantly, most solid tumors These localizations in the tumor are partially complete microvascular barriers (which (Separating the tumor tissue (interstitium)). This allows non- The injection dose reaching the intracellular reticulum target should be only about 1% to 7% (at most). And it is possible. Selected lymphomas and leukemias are larger than this vascular barrier. An exception to this rule is provided for serious corruption. However, most fixed General-purpose method still remains for morphological tumors and infections As a drug that efficiently passes through the microvascular barrier in a depot (controlled release) form You need to reach.   Such a drug form may cause normal vascular endothelium, organs, tissue cells To protect the drug from endothelial and cellular metabolism during transit, and And controlled therapeutic rate within target tissue and tissue trauma (including solid tumors) It is necessary to selectively make the drug bioavailable in.   Endothelial transport of active substances to small molecules (eg, glucose and insulin) Indicated. However, in some studies other than the active substance endothelial transport of the present inventors, For larger molecule (s) carried in the cargo format There is no indication of such transport. Current examples include polymer conjugates, and Non-covalent ion pair preparation of drug, and diagnostic agent having sulfurized glycosaminoglycan (The combined size is between about 8000 daltons and about 500 nanometers Transmigration through the endothelium occurs when sulfurized glycosaminoglycans and, in particular, Rumatan sulfate (which is a receptor or antigen (which is a disease-induced endothelium) More synthetic or synthesized at other sites but at the site of disease Accelerates by inclusion of (selectively binds to the receptor) (Ranney, Biochem. Pharmacology, Vol. 35, No. 7, 1063-1069). P. (1986)).   The present invention relates to improved novel compositions, carriers, agents and in vivo uses Describe the method. This method has improved selectivity, efficiency and efficiency at the site of disease. Provides a retraction mechanism and a kinetic-spatial profile You. In addition, the present invention provides for selective oligo drug localization in vivo, Improved selectivity, sensitivity of accumulation and action of unaffected (including solid tumors) Compositions, agents and drugs for degree, uptake mechanism and kinetic spatial profile And how to use it. The novel compositions are (a) uniquely non-covalently bonded and And (b) enhanced by physical stabilization. Other compositions are Prepared by covalent bonding. The bond is bound to the carrier (anionic or chemically acidic Contains saccharides, sulfatoids and glycosaminoglycans, typically And beneficially hydrophilic or substantially completely hydrophilic) Or with a chemically basic metal chelator. This active substance and key Carrier is also a combination of non-covalent, physical, and covalent means. Can be done by Non-covalent bonds include, but are not limited to, cationic or base Anionic or acidic saccharides with the appropriate ratio of the active drug to the metal chelate Carried out by means including the step of mixing with a carrier, whereby a strong, solution-like Dry and dry ion pair complexes and salts can be formed, respectively. this thing Is primarily gold to the anionic (acidic) group (s) of the acidic carrier. Based on the electrostatic binding of one or more cationic (basic) groups of the genus Chelator . Such bonds can include hydrogen bonds and physical factors (including, but not limited to, concentration, viscosity, And various drying means (including lyophilization).   Carrier materials useful in the present invention include, but are not limited to, natural and synthetic. , Native and modified, anionic or acidic saccharides, disacchara , Oligosaccharides, polysaccharides, and glycosaminoglycans (G AG) and, in particular, dermatan sulfate. A wide variety of additional biology Biocompatible, water-soluble and water-dispersible, anionic carrier materials can also be used This Is obvious to those skilled in the art. Water diffusion barrier, good initial biodistribution and multivalent site binding Oligomeric and polymeric, hydrophobic and Substantially completely hydrophilic carrier substances are used for tumors, cardiovascular infarcts and other types Among suitable carriers for drugs to be used to treat local diseases of the Included. However, amphoteric and hydrophobic carriers may be required for certain therapeutic applications. It will be clear to the skilled person that it may be suitable. In the present invention, the most useful drugs and And metal chelators include cationic groups for binding to carriers, and basic Containing a group and a basic amine group, and either directly or indirectly And those that are effective in treating local disease states. This is metal and gold Cleanup of genus ions, transition elements and ions, and lanthanide series elements and ions Includes Clean with cationic, basic and amine containing chelators Virtually any single atomic element or ion that can be The possibilities are also clear to the skilled person.   For the purposes of the present invention, cationic or basic metal chelators are: And can be further distinguished from metal ion complexes: cationic or Basic metal chelators are organic covalent cross-linkable ligand molecules (which are single Which may partially or completely surround the metal atom or ion), The resulting formation constant of the chelator for the appropriate metal or ion is at least , About 10¹⁴It is. A chelator can further be a chelator or a cationic or base That imparts properties and contains, but is not limited to, amine (s) The functional group (s) is completely or essentially completely electrostatic or Positively charged or protonated at the typical pH used for the agent A case is defined as cationic or basic. Formulation pH is dependent on vertebrate mammals. It is characteristically chosen to define the range of physiological pH present. This is Non-limiting examples include a pH in the range of 5 to 8. Amine is a metal Containing primary, secondary, tertiary or quaternary amines or combinations thereof May be included. As herein and as specified, the hydrophilic carrier is used in pharmaceutical formulations. Water-soluble under the conditions used for partitioning into the aqueous phase of a water-organic solvent mixture Or as a substance forming a translucent aqueous solution, complex, aggregate or particle dispersion Definition Is done. The carrier furthermore, whether it is completely or almost completely nucleophilic Or its functional group (s) are cationic, basic, or amine gold Can interact with the genus Chelator and completely or almost at the pH used for the formulation. When fully negatively charged, anionized or ionized, anionic or Is defined as acidic. Such anionic and acidic groups include Although not particularly limited, sulfates, phosphates and carboxyls on a carrier may be used. Sylate, or a combination thereof.   Novel pharmaceutical compositions include, but are not limited to, cationic or basic, representative Basically, a basic amine metal chelator active substance, or a metal chelator active substance ( Chelating metals or metal ions). here, These active substances may be natural or synthetic carriers, including but not limited to hydrophilic, On, or acidic, natural or synthetic, native, modified, derivatized And fragmented, anionic or acidic saccharides, Gosaccharides, polysaccharides, sulfatoids, and glycosaminoglis Anionic and acidic carriers, including can (GAG) Combine.   Anionic and acidic saccharide and glycosaminoglycan carriers are , Glucose, glucuronic acid, iduronic acid, glucosamine, galactose, glass Cutsamine, xylose, mannose, fucose, sialic acid, pentose and Other naturally occurring, semi-synthetic or synthetic monosaccharides or their chemistry Derivatives (which are amine, sulfate, carboxylate, sialyl, phos Monomer units, including fate, hydroxyl or other side groups) May be included. Glycosaminoglucan (GAG) is a carbohydrate moiety on the cell surface and Consisting essentially of tissue matrix proteoglycans. These are chemically separated Methods and extraction methods (and, in certain cases, enzymatic methods, see Lindahl et al., (1978), herein). Derived from naturally occurring proteoglycans Is done. These include, but are not limited to, the following types: Heparin, heparan sulfate, dermatan sulfate, chondroitin-4-sulfate, chondroline Itine-6-sulfate, keratan sulfate, syndecan, and hyaluronate , And over-sulfated, hyper-sulfated, and Other chemical derivatives are mentioned (further described below).   Strongly acidic, sulfated glycosaminoglycans include all of the classes listed immediately above. But excludes hyaluronate. This is due to the weaker acidic carboxyl It has only a sulfate group and does not contain a sulfate group. Natural sources of glycosaminoglycans Sources include, but are not limited to, porcine and bovine intestinal mucosa, lung, spleen, pancreas, and And various other solid and parenchymal organs and tissues.   Sulfatoids mainly (but not exclusively) include bacteria and A second class of sulfated saccharide materials derived from non-mammalian sources includes Can be Sulfatoids typically have shorter chain lengths than glucosaminoglycans. Although of low molecular weight, it can be modified synthetically and can give: Long chain length, (b) increased sulfation per unit saccharide, (c) various other chemical side chains Group, or (d) the desired ligand binding properties, and site-selective binding in vivo, Other properties that favor the uptake and accumulation characteristics (s). Sucrose and And other short oligosaccharides can be obtained from natural and synthetic sources.   These oligosaccharides include carboxylate, phosphate, sulfate Disaccharides using a salt or a silyl side group or a combination thereof With a substitution ratio of up to about 8 anionic or acidic substituents per unit, Can be made anionic or acidic by enzymatic derivatization. Repair Decorated glycosaminoglycans can be any of the natural glycosaminoglycans described above. Can be derived from ip and sources and include the following (1) and (2): (1) Glycosaminoglycan fragments (standard ion exchange separation and solvent fractionation methods) Has a shorter chain length than the parent material as directly isolated from natural sources (Further defined as glycosaminoglycans); (2) chemically modified to Glycosaminoglycan having reduced anticoagulant activity. This allows for "non-anticoagulant" (NAC A) GAG is obtained, which is not exclusive, but typically comprises (a) periodate oxidation, Then borohydride reduction; (b) partial or complete desulfation; and (c) non-covalent , Divalent or trivalent counterion salts (primarily, but not limited to, more acidic Calcium, magnesium, magnesium N Formation, including salts with cancer, iron, gadolinium and aluminum ions) Prepared by   For the purposes of the present invention, specific classes of such solution complexes and salts include: As described above, acidic saccharides or glycosaminoglycan carriers Or a sulfate group and a metal chelator or a metal chelator containing a metal. Electrostatic association or ion with basic or cationic group (s) Strong complexes and salts formed by pair association are included. Derivatized acidic filter Callides and glycosaminoglycans are typically species on the saccharide unit. It is prepared by derivatizing various chemical side groups at various sites. This is This can be done by chemical or enzymatic means.   Enzymatic means are used in certain cases where highly selective derivatization is desired. Obtained Chemical and enzymatic derivatives include, but are not limited to, those derivatized by And acid saccharides and glycosaminoglycans: (1) (a) carbo Esterification of a xylate group, (b) a hydroxyl group, and (c) a sulfate group; (2 ) Hypersulfation by non-selective chemical or selective enzymatic means; (3) acetyl And (4) various other ligand derivatives, including but not limited to (a) sialyl side groups (B) the addition of fucosyl side groups, and (c) various carbodiimides, anhydrides and And treatment with isothiocyanate bridging groups, and (d) attachment of various other ligands Addition).   The sulfate and sialyl side groups, if present, are And any of the glycosaminoglycan monomers (Lindahl et al., (1978), incorporated herein by reference). Certain resulting derivatized acidic saccharides and glycosaminoglycans are anticoagulant. The desired activity of solid activity, site localization pattern, clearance, and other biological properties May have modifications. Between certain classes of glycosaminoglycans and biological properties One example of this relationship is the natural sulfate / carboxylate ratio, which is preferred. Preferably in the range 0.7: 1 to 1.8: 1, more preferably between 0.9: 1 and 1.5: 1, And dermatan sulphate, typically having a 1: 1 ratio, binds to normal endothelial cells Is relatively low, avoiding the displacement of endogenous heparan sulfate from endothelial cell surfaces, Higher selectivity for induced endothelium at sites (including thrombi) And has rapid plasma clearance (essentially by the renal pathway) While higher sulfate / cal between 2: 1 and 3.7: 1 was reported. Heparin with a boxylate ratio and oversulfated dermatan sulfate are And relatively higher binding to both induced endothelium and endogenous endothelium. Substitutes relatively more paran sulfate and purifies more slowly than dermatan (Boneu et al., (1992), incorporated herein by reference). Is).   As newly described and used in the present invention, glycosaminoglycans The dermatan sulfate class of cans, and in particular a new specific class of dermatan sulfate (Which contain selectively oversulfated oligosaccharide sequences) The active substance associated with or bound to the active substance (ie, dermatan sulfate-active (DS-active substance) as a carrier substance, combined with extremely low toxicity It also has the unique benefit of higher potency. This is its (a) relative Low sulfate / carboxylate ratio (which is between 0.7: 1 and 1.8: 1) Range, most preferably between 0.9: 1 and 1.5: 1, and most representative (B) very low anticoagulant activity (very low factor Xa and U (For SP heparin activity and negligible binding to antithrombin III); (c) Very low or non-existent platelet aggregation and, for this reason, thrombocytopenia Conductive properties (which are less than about 45,000 daltons (preferably less than about 25,000 daltons) Relatively low SO combined with modal molecular weight_Three-/ COO- ratio); (d ) Essentially complete absence of metabolism in vivo; and (e) extremely rapid blood For medium and body clearance (all of which are described further below) . These properties allow for bleeding, metabolism, and in vivo in normal tissues and organs. Extreme in vivo safety profile with no residua Become higher. These properties and their resulting safety profile Class of glycosaminoglycans (GAGs) and other classes of acidic saccharides, Oligosaccharides, polysaccharides and sulfatoid substances (these To make up acidic and anionic saccharide substances) from Derma Tansulfate is clearly distinguished, and these are the other Unique and surprisingly superior to other classes of acidic and anionic saccharides Provide unexpected and unexpected benefits. Most particularly, dermatan sulfate is 2: 1 And SO in the range between 3.7: 1_Three-/ COO- ratio and IO greater than or equal to 10% C content (by weight-indicating these very high sulfate contents) Glycosaminoglycans surpass these surprising and predictable polysulfates Show no benefit. And most specifically, this new specific class of dermata Sulfuric acid (as described in detail below) And selectively enriched throughout the chain lengthas a wholeOversulfation or Does not include polysulphated molecules (the latter having a SO2 of greater than or equal to 2.0: 1)_Three-/ CO Characterized by an O-ratio and a sulfur content greater than or equal to 10%), The complementary, selectively persulfated males of these new specific dermatan sulfates The rigosaccharide sequence allows endothelium and tissue at disease sites (including tumors) Strongly binds to selectively induced receptors on matrix and target cells Have a further surprising and unexpected benefit. For this reason More, these new special dermatan sulfates are moderately low in theseAs a whole hand SO_Three ^-Extremely low cells based on / COO-ratio and sulfation and its associated To be expected based on toxicity and systemic toxicity properties and side effect profiles And surprising and unexpected stronger site localization and site targeting efficacy Is shown.   In certain cases unique to the present invention, acidic saccharides and glycosaminog Derivatization of the lican carrier can be achieved by the basic metal chelator itself. Up The general classes of carriers described above are particularly suitable for the present invention, but have a wide variety of Additional, natural, derivatized, and modified carriers, and the like It will be apparent to one skilled in the art that physical formulations may be particularly suitable for various applications of the present invention. is there. In one representative example, the source and type of glycosaminoglycan , Its chain length, and the sulfate / carboxylate ratio for the following purposes: Can be optimized: (1) Combine different small and macromolecule diagnostic agents and drugs (2) carrier endothelium on diseased versus normal endothelium In (3) minimizing dose-related side effects; (4) carriers Clearance rate and clearance of and associated diagnostic and therapeutically active substances Optimizing the lance route.   Non-covalent formulation of active substance and carrier allows for covalent chemical conjugates It gives a significantly higher active substance-to-carrier ratio than a high active substance-to-carrier ratio. In the present invention, non-covalent binding is a minimum of 15% by weight of active substance [active Substance / (active substance + carrier), w / w]; typically greater than about 30% (w / w); preferred At least about 50% (w / w); and often between about 70-99% (w / w) I can. Covalent bonds characterize the percent active substance by (a) (B) Peptides containing antibodies, smaller than about 12% for small and polymer carriers Less than about 7% for antibodies and protein carriers, and (c) antibody fragmentation. About 0% 5-2. Limit to less than 0%. This restriction applies to drug formulations and Functionalities Available on Carrier Molecules Useful in In Vivo and In Vivo Site Localization Based on the number of   Low substitution covalent active agent-carrier drug compositions are typically Non-covalent activity that can be useful for certain in vivo applications and yet has a high degree of substitution The substance-carrier drug composition is typically compatible with a wider range of other in vivo applications. It will be apparent to those skilled in the art that it may be useful. In general, but not exclusive However, non-covalent drugs are specialized for most diagnostic imaging and most therapeutic applications. Where high systemic and site-localized doses are required, And rapid clearance of the delocalized fraction of the administered drug is Plasma Clearan to maximize last and minimize systemic drug toxicity It is desirable to accelerate the speed and achieve low background levels.   These properties of the formulations of the present invention combine with prior art, non-selective and covalent Represents a further substantial improvement of the active substance-carrier drug obtained. The resulting drug (A) Site selection, including cardiovascular disease with tumor, infection and acute endothelial induction Selective drug localization; (b) MRI contrast and spectral enhancement, ultrasound contrast Enhancement, and X-ray contrast enhancement, where relatively high doses are preferred and May be required; (c) nuclear medicine or radionuclear imaging and therapy, where non-standard Increased clearance of targeted doses may be preferred or required: and (d) features Constant or high doses of extended release or sustained effect treatments are preferred or needed It can be widely useful where possible. Such therapeutic agents include (a) acute vascular ischemia , Acute infarction, acute vascular injury, shock, hypotension, restenosis, tumor and tumor blood Tube formation and soft tissue cells or other pathological growth; and (b) the following classes of diseases: Affected: Blood vessels, soft tissues, mesenchyme, endothelium, smooth muscle, striated muscle, adventitia, immunity, inflammation, bacteria, bacteria Treatment of species, viruses, degeneration, neoplasms, genetics and enzymes Including but not limited to drugs that are useful in medical and medical indications.   MRI contrast enhancement and pharmacotherapy are important indications, High revenue loading and controlled release of active agents is important in addition to site-selective localization. A particular advantage (see below).   For the purposes of the present invention, potential chelating agents that are generally useful for transchelation at the site of disease Metal ions are iron, manganese, chromium, copper, aluminum, nickel Gallium, indium, gadolinium, erbium, europium, disp Divalent and trivalent cations selected from the group consisting of rosium and holmium May be included. Radiation for nuclear imaging and compositions and uses, and radiation Chelating metal ions generally useful in radiotherapy compositions and uses are phosphorus, sulfur , Gallium, iodine, germanium, cobalt, calcium, rubidium, Thorium, technetium, ruthenium, rhenium, indium, tin, iridium Selected from the group consisting of, platinum, thallium, strontium and samarium It may include metal. Metal ions useful in neutron capture radiation therapy are boron and boron. And others with large nuclear cross sections. Ultrasonic contrast composition and Metal ions useful in X-ray and X-ray contrast compositions and their use are those in which It may include any of the metal ions listed above, provided that appropriate site concentrations are achieved. And, in particular, may comprise a metal ion having an atomic number at least equal to the atomic number of iron You.   For the purpose of the present invention, treatment in the chelation of iron, copper or both inside the body The agents for the composition and use include these metals ((1) they are typically Required for formation, or (2) they cause and amplify local tissue damage [Levine (1993), hereby incorporated by reference)). Carrier with a base metal chelator in one or both of the following forms: Including: (a) chelator without carrier + metal ion; and (b) carrier + metal The individual base metal chelators of this composition should be chelated by ion exchange. Added to the composition with a formation constant lower or equal to the formation constant of the metal inside the body. A chelating agent having a chelated metal ion (see below). Such weakly chelated metal ions of the composition include calcium, manganese, Magnesium, chromium, copper, zinc, nickel, iron, gallium, indium, From aluminum, cobalt, gadolinium or other exchangeable ions. Metal ions selected from the group consisting of: Compositions for other therapeutic uses Useful metal ions for inclusion in magnesium, manganese, chromium, zinc and And divalent and selected from the group consisting of calcium, iron, copper and aluminum. And trivalent cations. Each of the above metal ions is identified above May be used in combination with a base metal chelator for other indications than the indicated indication. And that, under certain conditions, metal ions other than those listed above It will be apparent to those skilled in the art that the listed uses and indications may be useful.   The compositions described in the present invention provide surprising and unexpected improvements in performance and use. And they include:   (1) Active substances and caps retained during in vitro dialysis and in vivo targeting High meeting with Leah;   (2) Selective binding of active substance and carrier to induced endothelium at disease site If;   (3) After intravenous administration, rapid selective endothelial binding, disease-induced endothelium (histologically Encapsulation and extracellular migration of carriers and metal chelators (including porous endothelium) Very rapid (2-7 minutes) localization at the site of the disease due to emergence;   (4) widespread uptake across diseased tissue sites;   (5) Sustained retention (several hours to several days) combined with:   (6) rapid plasma clearance of non-targeted fractions (min);   (7) Moderately slower to plasma, giving prolonged disease site retention, polymer reverse Diffusion rate;   (8) Solid tumors or acute blood vessels in body sites, and in brain and central nervous system sites And myocardial infarction, including small tumor metastases in the liver, lungs and other organ sites, Ability to selectively treat and image with substantially improved selectivity and sensitivity .   The enhancement of diagnostics and drugs can be caused by a number of mechanisms, One is: 1. Effective targeting of disease sites to tissue; 2. Stabilization during both storage and plasma passage; 3. Prolongation at the disease site, giving a significantly increased area under the curve at the tissue site Retained; 4. Rapid clearance of non-targeted fractions, thereby providing background in imaging applications Signal and reduce normal organ exposure and systemic toxins in therapeutic applications Reduce sex. Five further significant advantages of the compositions and uses of the present invention are:   1. Simple formulation of active substance and carrier;   2. Stabilization of diagnostic and therapeutically active substances during storage and transit of plasma;   3. Rapid site localization and persistent site retention;   4. Rapid clearance of the non-targeted fraction;   5. Low-toxic carbohydrate carriers and glycosaminoglycans from natural sources Carrier availability and, if necessary, modification or Derivatization.   Acidic or anionic saccharides and glycosaminoglycans are used in vivo. Has a unique mechanism of site localization and retention. These are the underlying tissue diseases. It binds to endothelial determinants of the body that are selectively induced on the microvascular barrier by disease. Department Earlier approaches to targeting sites have turned to antigenic determinants. However, these decisions Determinants typically traverse an isolated site in tissue, in other words, across the endothelial barrier, And because it is located in the bloodstream and not on its endothelial surface, Carriers and drugs injected into the bloodstream recognize these areas of the target antigen, And there is no effective means for localization. In other words, the application Recognizes vascular access codes required for effective extracellular translocation at disease sites Ignored the main problem of improper carrier distribution arising from not doing so. Follow Thus, these carriers contain their binding actives in effective concentrations at the relevant tissue sites. Could not be filled effectively.   Glycosaminoglycans, dermatan sulfate and a new specialty dermatan sulfate Acidic or anionic saccharides, which initially contain complementary By binding to the receptor, it is localized at the target site, Induces a very rapid (approximately 3 minutes) extracellular transmigration of the active Penetrates widely into the ricks, the site of the disease (tumor-typically within the tumor matrix) Are several hundred micrometres from irregularly spaced and perfused microvessels. (Including tumors at a distance to the mediator) to select a carrier-drug accumulation reservoir. Alternatively, and thirdly, complementary receptors on the final target cells (including tumor cells). GAG-active substances (including DS-active substances) to induced tumor cells Leads to internalization (see example below). (Listed immediately before and further below) A new special class of dermatan sulfate (described in more detail in Complementary binding via a highly sulfated saccharide sequence that is highly concentrated These sequences have at least about 220 U / mg of concentrated heparin Correlate with cofactor II activity, and these sequences are Positively charged, selectively induced on ricks and target cells (including in tumors) To a cationic and / or structurally complementary receptor or lectin Seems to combine. For the novel dermatan sulfate, these binding and Target function and capacity are (as described elsewhere herein) overall high sulfation / It occurs without either polysulfation or the inherent toxicity and safety disadvantages.   The biological address of the disease site can be found in a manner similar to that of the postal address. Here, effective carrier substances are (1) first induced by tissue disorders at the base of the body. (2) Then recognize the extracellular tissue matrix A local and effective dose of diagnostic and therapeutic active Extracellular transmigration and packing; and (3) Finally, the target cell and antigen And bind to address. The earlier approach to site delivery is the first Recognize "street" addresses without recognizing "state" and "city" addresses. Was trying.   The acidic saccharide and sulfated glycosaminoglycan systems are used for the previous antigen-recognition The reason that they work substantially better than approaches is that they Newly induced signal used to attract and target leukocyte cells to It is to recognize. When the disease strikes a local site, it is within the tissue matrix At the endothelium-blood interface, it initiates a cascade of local mediators, which Disease and immune cells need signals within tissue sites to signal blood cells and the central body system send. These mediators include cytokines, chemoattractants, cytotoxic Cells, induced cell-surface adhesion, selectins and integrins, and various Tissue-derived and blood-borne, soluble and cell-surface procoagulants No. Leukocyte accumulation starts within minutes and the nature, severity and persistence of local disease And the generation of continuous tissue mediators and transendothelial signals And last for several days to several weeks.   As currently reported and reviewed in detail [Ranney (1990); Ranne y (1992); Bevilaqua et al. (1993); Bevilaqua et al. (1993); Travis (1993); Sharon et al. 93), all of which are incorporated herein by reference], tumors, infarcts, infections, inflammations Sexual disease, vascular abnormalities, and other focal disorders are associated with resident macrophages and local From the tissue matrix, such host mediators, or cytokines Characteristically induces the release of In certain diseases, bacterial lipopolysaccharide (LPS), viral R Exogenous mediators such as NA, and tumor-derived inducers including EMAP II Sir, and chemical inducers can also be released. More mediators theory Still to be clarified, but main mediators are defined and include: : Interleukin 1 (IL-1), tumor necrosis factor (TNF), vascular endothelial growth factor / vascular penetration Sex factor (VEGF / VPF), transforming growth factor β (TGF-β), lipopolysaccharide (LPS) , Single- and double-stranded nucleotides, various interferons, monocyte chemoattraction Protein (MCP), interleukin 8 (IL-8), interleukin 3 (IL-3), Interleukin 6 (IL-6), a tumor-derived inducer (as described above) and chemical Attractant peptides, various prostaglandins and thromboxanes. Described earlier Specific mediators induce local production and release of metalloproteinases, And this is in turn the intact and partially truncated integrin, RDGS Peptides, laminin, collagen, fibronectin, and glycosaminoglyc Exposing potential tissue binding sites, including the cell-surface core protein components of can You.   Cytokines including VEGF / VPF and monocyte chemoattractant protein (MCP); Tissue metalloproteinases and proteolytic tissue matrix flags Cells directly induce local endothelium and circulating leukocytes, including neutrophils, monocytes and lymphocytes. Become adherent to spherical cells. The induced endothelial adhesion molecule (adhesin) is Including: P-selectin (gmp-140), E-selectin (ELAM-1), intracellular cell adhesion molecule ( ICAM-1), vascular cell adhesion molecule (VCAM-1), inducible cell adhesion molecule, (INCAM-110), Invirbrand factor (vWF, factor VIII antigen) (these individual types of endothelial See below for disease states that activate syn). Further cascade Become activated and indirectly amplify endothelial adhesion. These are (1) Solid factors, especially fibronectin, tissue factor, thrombin, fibrinogen, Fibrin and its split products, especially fibronectin split products and fibrin Brinopeptide A; (2) Platelet-derived factor: Platelet activating factor (PAF), glycoprotein IIb / IIIa complex; (3) white blood cells containing VLA-4 (very late antigen 4) (a) L-selectin , And (b) integrins; and (4) many complement factors.   Prior pathological processes and signals may be directly or indirectly: Contains acidic saccharides (AC) and glucosaminoglycans (GAG) Involved in binding and site localization (see below for potential tissue binding sites Note that are called "GAG" and "AC").   1. Local tissue disease, as described above, local cytokines and mediators Is induced. In particular, the cytokines vascular endothelial growth factor / vascular permeability factor (VEGF / VPF) selectively by many or most tumors of human and animal origin Derived [Senger et al. (1994), incorporated herein by reference], and specific 34-42 kDa heparin binding that acts directly on endothelial cells via endothelial receptors Be a compatible and GAG binding glycoprotein [Jakeman et al. (1993), incorporated herein by reference. Which has been recently reported to cause endothelial activation and G Induce additional new endothelial receptors capable of binding AG (see below). VEGF / VPF has very high potential for endothelial vs. fibroblasts and other cell types. Are selective, chemically basic growth factors [Senger et al. (1994); Nicosia et al. 4), incorporated herein by reference]. This is the case for many or most humans And long-term endothelial angiogenesis in animal tumors and AIDS-associated Kaposi's sarcoma It appears to be a key growth factor to stimulate (Connolly et al. (1989); Weindel et al. (199) 2), both incorporated herein by reference). In certain instances, VEGF / VPF is More temporary and anatomically restricted vessels of wound healing and vascular restenosis May be important for the formation process [Senger et al. (1994); Miller et al. (1994); Nicosia et al. (1 994); Berse et al. (1992), all of which are incorporated herein by reference]. VEGF / VPF And platelet-derived growth factor PDGF-BB have significant angiogenic potential in vitro The only species in the group of basic GAG-binding growth factors, namely the in vivo cofactors It has recently been reported to be a species that is directly active in the absence [Nicosia et al. (1994) , Incorporated herein by reference]. The effect of VEGF / VPF is on the outer surface of this molecule. Inhibited by antibodies to specific peptides above [Sioussat et al. (1993); And, importantly, such inhibition is in vivo. Inhibits the growth of animal tumors [Kim et al. (1993), incorporated herein by reference]. . Thus, both VEGF / VPF are present in tumors and potentially in other pathological lesions. To provide and target receptor targets for binding of GAG carrier substances You.   2. These cytokines and mediators include VEGF / VPF, MCP (Yamashir o et al., 1994) and induce histochemical attractants, including IL-8, which bind GAG / AC It has been reported that arginine-rich, 8Kd heparin-binding protein Including the family [Huber et al. (1991), incorporated herein by reference];   3. No. above 1 cytokine and mediator induces local endothelium, A reported binding determinant for GAG / AC [Bevilaqua et al. (1993); Bevilacqua et al. (1993)], P-selectin, vascular cell adhesion molecule (VCAM-1), inducible cell adhesion molecule (INCA M-110), and von Willebrand factor (vWF, factor VIII antigen); Selectins have been reported to bind to GAGs [Bevilacqua et al. (1993)];   4. Integrins, including but not limited to VLA-4, are involved in a variety of disease processes Is induced on circulating white blood cells, including lymphocytes (see below); VLA-4 has a distinct binding site on the (induced) endothelial selectin VCAM-1 (see above). No. 3); abundant in plasma and also obtained from tissue sites The resulting fibronectin has distinct and discrete binding sites on VLA-4 [Elic es et al. (1990)]; fibronectin has a specific binding site for GAG / AC. [Bevilaqua et al. (1993)] indicate that these amplification steps are powerful for site localization of GAG / AC. Provide a mechanism consisting of:   5. Chemical inducers, MCP and IL-8, lymphocyte integrins, VLA-4, platelet factor Offspring, PAF and coagulation factors, thrombin, fibronectin and others Spread from tissue and blood, bind to the induced endothelial selectin, and Amplifies adhesion and activation at early endothelial P-selectin sites for GAG / AC [Elicse et al. (1990); Lorant et al. (1993)];   6. Tissue metalloproteinases become activated and are activated Exposing new binding sites for GAG / AC in the underlying tissue of the damaged endothelium. this These new tissue binding sites include the following [Ranney (1990); Ranney (1992); Travis (19). 93); Bevilaqua et al. (1993)]:         a. Fibronectin fragments;         b. Collagen fragments;         c. Laminin fragment;         d. RGDS peptide;         e. The exposed core protein of GAG;   7. White blood cells are attracted to this site, become activated, and Release proteolytic enzymes, thereby producing No. Amplify 6, and tissue Increase the exposure of the binding site for GAG / AC in the matrix.   8. GAG / AC carriers are listed in No. Induction and exposed decisions listed in 1-7 At the lectin-carbohydrate level, which selectively binds to groups and is characteristic of leukocyte cell adhesion, Confer an immune-type localization that is specific for the induced binding site (lectin);   9. When the carrier substance has a multivalent bond to the target binding substance (e.g., Carrier is acidic oligosaccharide or polysaccharide or acidic glycosa Multivalent bonds on the endothelial surface, including carrier and binding Induces rapid extracellular transmigration of activity and binding of antibodies, liposomes, and monovalent For filling achieved by substances (such as hormones and monovalent-linked sugars) Resulting in substantially increased packing of the tissue matrix;   Ten. GAG / AC adhesion to induced and exposed tissue binding sites Reduces plasma back-diffusion of these binding actives, thereby maintaining persistence within the tissue site Give retention;   11． Diagnosis or controlled release of drug activity from carriers containing GAG / AC within disease sites , Which results in a targeted controlled release;   12． Tumor cells, microbial targets and damaged cell targets within the tissue site are GAG / AC Selectively capture bound diagnostic or drug-active substances, respectively, based on: M: Induced tumor anion transport pathway, microbial binding site for GAG / AC, and tamper Cell-surface core proteins exposed by cytolysis [Ranney 07 / 880,660, 07/80 3,595 and 07 / 642,033] --- Uptake, Cr4S uptake by prostate adenocarcinoma [Kjellen et al. (1977)]. 13. If the carriers are hydrophilic or essentially completely hydrophilic, these carriers Separates the binding active from the typically irregularly spaced microvessels of the tumor. Deep into the tumor mass and throughout the tumor mass, even at the isolated histological site ( Permeate). And also the border of normal tissue around each lesion of the disease (typically (Including edges about 30-75 μm thick). But like this The carrier and / or the active substance (diagnostic or therapeutic) associated therewith, Undergo selective uptake (internalization) by abnormal cells in the tissue site, And preferentially avoids uptake by normal cells within the site, thereby: a. For diagnostic imaging applications: border between tumor or infarction and surrounding normal tissue Very sharp resolution of b. For therapeutic applications:   (1) protection against the spread of disease at the edges;   (2) inside and adjacent to the disease site from uptake of cytotoxic drugs Relative protection of normal cells give. 14． In the case of a hydrophilic carrier (including but not limited to GAG / AC), the active The quality non-targeted fraction is rapidly and non-toxically purified, thereby: a. In imaging applications, background signal intensity; b. For all applications:   (1) normal organ exposure; and   (2) Systemic side effects Is minimized.   In view of the above overview, tumor-selective GAG binding cytokines, VEGF / VPF and And MCP are currently known to exist in three histological locations: Tumor cell surface, tumor extracellular matrix, and localized tumor angiogenic endothelium. therefore , These cytokines are responsible for all three major tumor sites, namely tumor endothelium GAG-drugs in the tumor extracellular matrix and the tumor cells themselves Of the receptor. These cytokines are selectively located on tumor endothelium In Intravenously administered GAG-drugs selectively bind to tumor microvessels. GAG-drugs can pass through VEGF / VPF- "permeabilized" endothelium Thus, it is possible to selectively overflow very quickly (about 3 minutes). The following points Note that such "penetration enabled" has actually recently (a) expanded significantly (Larger than the standard 120 nm Palade vesicles that characterize normal endothelium) and (normal Rapid transport by vesicular endosomes with significantly increased numbers (than vascular endothelium) [Seng er et al., (1993), incorporated herein by reference]; and (b) True microfiltration porosity is assessed by macromolecular and particulate markers As such, it has been shown to include anatomically non-porous vascular endothelium. below As shown in specific examples, VEGF / VPF and MCP cytokines on tumor cell surfaces The presence of in could explain the selective tumor cell internalization of the GAG-drug. The important thing is No. of these cytokines and GAG binding peptides 6 (above) is tumor matrix Present in large extracellular volumes of GAG-binding agents (including metal chelates) Partly describes a large tumor tissue reservoir (MRI contrast). (See Examples below). Such drugs into the bloodstream Due to the relatively slow backdiffusion (about 7 hours), such extracellular The presence of tissue-matrix receptors is further enhanced. The important thing is: (1 ) Prolonged tumor retention of Gag-drug as extracellular reservoir (depot) (B) that some of this extracellular drug is internalized into tumor cells; and (c) The combination of very rapid clearance of the targeting moiety from the blood and the body The combination provides in vivo imaging (including MRI contrast enhancement) and cure The following surprising and unexpected benefits are provided for treatment: (a) Enhanced tumor Tumor selectivity; (b) prolonged, high, "area under the curve" (AUC) in tumor; (c) blood Short and low AUC; (d) Minimization of local and systemic toxicity. In addition, Including (A) cytokines and mediators; and (B) selecti Selections, integrins, and adhesins report on these tumors Has been reported to be induced by various disease symptoms in addition to those reported [B evilaqua et al. (1993)]. Representative non-oncological induction is also seen below. A. Cytokines and mediators.   1. MCP: experimental autoimmune encephalomyelitis in mice [Ransohoff et al. (1993)];   2. IL-8: Angiogenesis: [Strieter et al., (1992)];   3. PAF: Reperfused ischemic heart [Montrucchio et al., (1993)]. B. Selectins, integrins, and adhesins.   1. ELAM-1:     a. Hepatic portal endothelium in acute and chronic inflammation and allograft rejection [Steinhoff et al. (1993)];     b. Active inflammatory processes, including acute appendicitis [Rice et al., (1992)].   2. VCAM-1:     a. Monkey AIDS encephalitis [Sasseville et al., (1992)].     b. Allograft rejection of liver and pancreas [Bacchi et al., (1993)].   3. INCAM-110: Chronic inflammatory diseases, including sarcoidosis [Rice et al., (1991) ].   4. Integrin, β1 subunit cell adhesion receptor: inflammatory synovial ream Yui [Nikkari et al. (1993)].   Diagnostic use for a broad category of focal tissue disorders and many specific types The carrier and active substance of the invention may be directed both for Is clear from the above. These include tumors, cardiovascular diseases, inflammatory diseases, Bacterial and viral (AIDS) infections, CNS degenerative disorders, and allogeneic transplants Includes partial rejection. Carriers disclosed in the present invention for many other disease states It will also be apparent to those skilled in the art that   Glycosaminoglycan (GAG) site selectivity is more likely than leukocyte rather than antibody targeting It appears to mimic the targeted level of the immune system. Antibodies have very high specificity As such, antibodies characteristically remove major subregions of disease lesions (references). Typically, as many as 60% of the tumor cells are unbound). Recently, endothelial P-selectin Of GAG binding determinants has been identified as sialyl Lewis x Was. Others are in the process of identification. Notably, sialyl Lewis x Specific and available nonvalent oligosaccharides have two critical Problems are:   1. These are extremely expensive substances and can only be obtained by synthetic or semi-synthetic means. Available and therefore cost-effective;   2. They do not bind effectively to disease sites under in vivo conditions. This is this A monomeric binding substance that displaces the endogenous interfering substance previously bound to these sites. It seems to be due to their inability to do so.   The range of GAG specificity is relatively wide, and the GAG binding site is Tricks and being abundant on cells have two distinct advantages Do:   1. GAGs cover a wider range of tumors and diseases than can be targeted by antibodies. (Antibodies are typically of one type of histological type. Target only (even within a given class of tumors) and therefore the antibody Not typically effective from both a medical and cost / development perspective );   2. GAGs are used for longer time intervals (from early onset of disease to progression and regression) It is thrown in effectively throughout.   Despite wider targeting specificity of GAGs than antibodies, its preferred clearing Avoiding uptake by cell and normal cells, even if more than one species The disease site of the target is targeted accumulation of the diagnostic agent / drug within its extracellular matrix If possible, systemic and local toxicity is reduced.   The polymer properties and multivalent binding properties of GAGs are both It is very important for optimal site localization of the object. The molecular weight of GAG is usually about 8,000 ~ 45,000 MW, preferably 10,000-23,000 MW, and more preferably 13,000-19,0 00 MW is important for:   1. Limiting the initial biodistribution of diagnostics / drugs to the plasma compartment, Maximizing the amount of drug available for more site targeting;   2. Replacing endogenous interfering substances previously bound to the disease endothelium;   3. GAG-Active Incorporation of Diagnostics / Drugs into Underlying Tissue Matrix Inducing skin translocation;   4. Provides rapid clearance of added active substance and significantly reduced side effects To offer.Summary of the Invention   In all of the following descriptions, paramagnetic metal ion chelates and thereby The resulting images show drug localization at disease sites, including tumor sites And disease site levels, distribution, and residence time, and blood and Organ clearance patterns and kinetics (all of these are For the treatment of local sites of tumors, infections, cardiovascular diseases, and other diseases as described in (Including useful agents) targeting, localization, accumulation, cellular internalization, and blood and And may be useful in determining body clearance). Intend.   The present invention includes novel agents, which are cationic or chemically basic, Amphoteric or hydrophobic therapeutic agents (including peptides, polypeptides, and proteins ), And a metal chelator and metal ion chelate with a hydrophilic carrier. Anionic or chemically acidic saccharides, sulfatoids toid), and glycosaminoglycans. The present invention In embodiments, the agent also comprises a chelated metal and metal ion . The binding of the metal chelator to the carrier can be by covalent or non-covalent chemical or physical Stabilized by strategic means. In some embodiments, the novel non-covalent bond The composition provides an unparalleled high payload of the metal chelator to the carrier And provide the ratio. This is from about 15% by weight of metal chelator, 70% by weight % To 90% by weight of the metal chelator. concrete Embodiments include deferoxamine, ferrioxamine, iron-basic Porphine, iron-triethylenetetramine, gadolinium DTPA-lysine, gadolini N-methyl-1,3-propanediamine (N-MPD) -DTPA, gadolinium DOTA-lysine, And basic derivatives of gadolinium and porphyrin, polyfin, expanded (expanded) Porphyrin, Texaphyrin, and Saphirin (sap phyrin) as a basic or cationic metal chelator, Binds to a neutral or anionic carrier. One acidic or anionic carrier Ma Or higher acidic or anionic saccharides, Claus, pentosan polysulfate, dermatan sulfate, essence Selectively purified dermatan sulphate (sulfur content up to 9% (w / w) and selective Oversulfation of oligosaccharides), heparan Sulfate, bovine heparin, porcine heparin, non-anticoagulated heparin, and other natural and And modified acid saccharides and glycosaminoglycans.   Methods of contrast enhancement of magnetic resonance imaging (MRI) are described in certain embodiments of the invention. Yes, this is due to metal chelators and chemists during the transit of plasma in vivo. Very fast, metal-chelator composition based on a stable bond with the carrier Carrier-mediated site-selective in vivo localization and sustained site retention Confirmation allows site localization after intravenous administration. Quick and selective endothelial site Binding provides rapid extravasation to the underlying tissue site, accumulation at the site, and persistent site retention. And, at the same time, rapid clearance of non-localized fractions. By using the bright composition, selective MRI contrast enhancement of tumor and Demonstrated in cardiovascular infarction.   Mechanisms of selectivity, localization and cellular uptake, and sensitivity of MRI contrast A surprising and unexpected improvement in metal chelates with standard paramagnetic effects Is shown. A further advantage of the use of the compositions and methods of the present invention is that ), Which includes evidence of special histological staining, Site-selective endothelial binding, extravasation, tissue matrix accumulation, and cellular uptake mechanisms Confirmed. The effectiveness of selective localization and MRI imaging is also due to paramagnetic metal Occurs when the active substance is administered in a carrier-bound form, It is shown that the form does not occur.   In its most general embodiment, the present invention provides a chelator for metal ions. A chelator having a cationic group and an anion Bound to a hydrophilic hydrophilic carrier. In another embodiment, the metal ion Chelators with cationic groups are anionic due to non-covalent electrostatic bonding. Bonds with hydrophilic carrier. And, in another embodiment, the present invention provides a method of manufacturing A drug containing a basic chelator for the And has a covalent bond to an anionic hydrophilic carrier. Clean In certain embodiments of the invention, wherein the linker is not covalently attached to the carrier, the key The lator may be basic.   In one embodiment of the invention, the chelator for metal ions is an anion. Contains a chelator having a cationic group bonded to an on-hydrophilic carrier The drug may further contain a chelated metal ion, and especially if it is paramagnetic It may further contain metal ions. Non-covalently linked to the agent of the present invention, particularly the carrier. The combined agent of the invention comprising a chelator for metal ions is at least It can be further defined by about 15% by weight of the chelator. Preferably, the key The radiator should have at least about 10¹⁴Has a generation constant of   The agent of the present invention containing a metal ion is preferably iron, magnesium, chromium, Copper, nickel, gadolinium, erbium, europium, dysprosium, And holmium. In one embodiment, Therefore, the agent of the present invention comprises boron, magnesium, aluminum, gallium, gel Manium, zinc, cobalt, calcium, rubidium, yttrium, technetic U, ruthenium, rhenium, indium, iridium, platinum, thallium, summer Even a metal ion selected from the group consisting of lithium, tin, and strontium May be included. In certain embodiments, non-radioactive or radioactive phosphorus, sulfur, or The iodine can directly bind to the carrier (described later). Functionally with the listed metal ions Equivalent other metal ions may also be included, and are within the scope of the claims and It is understood to be within the spirit.   In one preferred embodiment of the invention, the medicament comprises a carrier, wherein the key Carriers are acidic saccharides, oligosaccharides, polysaccharides, Is a glycosaminoglycan. The carrier is also acidic glycosaminoglycan Or it may be a sulfatoid. In particular, carriers include heparin and desulfated heparin. Phosphorus (desulfated heparin), heparin bound to glycine, heparan sulfate, del Matan sulfate, essentially purified dermatan sulfate (with sulfur content up to 9% (w / w) And selective over-sulfation of oligosaccharides) Lonic acid, pentosan polysulfate, dextran sulfate, sulfated cyclodextrin, Alternatively, it may be, but is not limited to, sulfated sucrose.   In one embodiment of the invention, the chelator is a chelator for iron ions is there. Preferably, the chelator is hydroxamate, and And more preferably deferoxamine. In one embodiment, the metal ion Chelators combined with are ferrichrome, ferrioxamine, enterova Cutin, ferimicobacter, or ferrichrysin. Special In a preferred embodiment, the chelator is deferoxamine and the carrier Is heparin, or a heparin fragment, and the drug further comprises iron (III). In another embodiment, the chelator is deferoxamine, The carrier is dermatan sulfate or a dermatan sulfate fragment. Thus, the agent may further comprise chelated iron (III).   In certain embodiments, the present invention also relates to heparin, heparan sulfate, dermatan. Sulfuric acid, essentially purified dermatan sulfate (sulfur content up to 9% (w / w); And selective oversulfation of oligosaccharides) or Deferoxamine bound to a carrier selected from the group consisting of leutin sulfate And may further include metal ions. The agent of the present invention may also , A chelator that is a porphyrin, saphirin, or texaphyrin And this is furthermore a metal ion, and preferably an iron ion or gadolinium Mion may be included.   In a particularly preferred embodiment, the agent of the invention comprises 5,10,15,20-tetrakis (1- (Methyl-4-pyridyl) -21H, 23-porphine chelator, heparin cap Rear, and chelated iron ions. In some embodiments, the key The radiator can also be a polyaminocarboxylate or a macrocycle, and Preferably a basic or amine derivative diethylenetriaminetetraacetate 1,4,7,10-tetra of a basic or amine derivative Azacyclododecane-N, N ', N ", N"'-tetraacetate (DOTA). The present invention In some drugs, the carrier is also an endothelial determinant that is selectively induced at the disease site. May be further defined as being complementary to   In certain embodiments, the invention is derived from an induced magnetic resonance signal. An image-enhancing agent or a spectrum-enhancing agent for enhancing an image, wherein the agent Is 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide, N-ethoxy Bonyl-2-ethoxy-1,2-dihydroquinoline or carbonyldiimidazole Thus, it includes ferrioxamine covalently linked to heparin. Alternatively, the present invention , Spectral enhancement to enhance images obtained from induced magnetic resonance signals It is a strong drug that can be heparin, dermatan sulfate, or essentially purified. Dermatan sulfate (sulfur content up to 9% (w / w) and selective oligosa Gd (III) covalently linked to one of the Contains diethylenetriaminepentaacetate. Alternatively, the present invention provides An agent for image formation, which is a basic chelate for metal ions. And a chelated metal ion, the chelator comprising heparin , Desulfated heparin, heparin combined with glycine, heparan sulfate, dermatan Sulfuric acid, essentially purified dermatan sulfate (sulfur content up to 9% (w / w); And selective oligosaccharide oversulfation), hyaluronic acid , Pentosan polysulfate, dextran sulfate, sulfated cyclodextrin, or Non-covalent electrostatic bond with hydrophilic carrier selected from the group consisting of sulfated sucrose Therefore, they are connected. Agents for enhancing body imaging are preferably deferred. Loxamine, chelated Fe (III), and its associated deferoxamine A glycosaminoglycan carrier, and more preferably the glycosaminoglycan The can carrier is dermatan sulfate and / or Fe (III) is radiopharmaceutical ( radiopharmaceutical) metal ions, and most preferably the radiopharmaceutical Metal ions⁵⁹Iron or⁶⁷Gallium.   In another preferred embodiment, the present invention provides a method for enhancing body imaging. A drug, wherein the drug is diethylenetriaminepentaacetate-lysine or Is N-methyl-1,3-propanediamine DTPA, chelated Gd (III) and Glycosaminoglycosylated on Diethylenetriaminepentaacetate-lysine Contains can carriers. Alternatively, the present invention provides a method for enhancing body imaging. A drug comprising DOTA-lysine, chelated Gd (III), and The above 1,4,7,10-tetraazacyclododecane-N, N ', N' ', N' ''-tetraacetate- Contains a glycosaminoglycan carrier linked to lysine (DOTA-lysine). In certain embodiments, the present invention relates to dermatan sulfate, or up to 9% (w / w). Oversulfation of selective oligosaccharides with a sulfur content lfation) and non-covalent electrostatic binding to essentially purified dermatan sulfate Is a drug containing ferrioxamine linked by   In a further preferred embodiment, the present invention enhances body imaging An agent for MRI imaging and spectral shift, the agent comprising Is N-methyl-1,3-propanediamine-diethylenetriaminepentaacetate ( N-MPD-DTPA) containing chelated gadolinium (III) Is, or most preferably, bound by ion pairs or other non-covalent means Associated or associated, or preferably by covalent means. Cosaminoglycans, preferably dermatan sulfate, and most preferably new A special class of dermatan sulfate, and most preferably, selectively oversulfated A new special class of dermatan sulfate containing a modified oligosaccharide sequence Join.   Any of the agents of the invention as described in the above paragraphs or appended claims Should be buffered, saccharide-free to provide adequate osmotic strength for parenteral administration. As a combination with at least one of a salt, a sulfated saccharide, or a salt, and Lyophilization or lyophilization suitable for aqueous and aqueous solutions or aqueous regenerated products having the desired osmotic strength. Is further defined as a dry preparation, where the drug is sterile or sterile It is understood that this can be done.   Another embodiment of the present invention provides a method for imaging magnetic resonance images or spectra in vertebrates. This method involves adding a metal ion chelator to the animal. An effective amount of the agent of the present invention containing the carrier described above and a paramagnetic ion. Process. In particular, the present invention relates to images generated from induced magnetic resonance signals. A method for enhancing images in vivo, comprising the agent of the invention containing paramagnetic ions Administering to a subject an effective amount of the subject, exposing the subject to magnetic fields, and radio frequency pulses. The induced magnetic resonance signal to obtain a contrast effect. Obtaining step.   In another embodiment, the invention is a method of enhancing in vivo imaging. Then, an effective amount of the agent of the present invention containing a chelated metal ion is administered to a subject. Applying, exposing the body to ultrasound or X-rays, and obtaining a contrast effect Measuring the change in signal.   In another embodiment, the invention is a method of obtaining an image of a body in vivo, comprising: An effective amount of the agent of the present invention containing a metal ion whose metal ion is a radioisotope Administering to the subject and measuring the scintigraphic signal to obtain an image Step.   In another embodiment, the invention provides local and systemic toxicity and side effects. Effectively, quickly, and / or non-toxic for non-targeting to minimize While purifying the amount, the selective delivery, localization of the therapeutically active substance to the site of local disease, And compositions and methods for maintenance. Has blood vessels or other nearby pathways And induced vascular receptors, adhesins, or as described herein. If it has any other form of signal that can be recognized by the carrier material being These therapeutically active substances and treatment methods can be used for any type and location of local disease site. To get. In particular, such actives for the treatment of tumors include: But not limited to: doxorubicin, adriamycin, taxol, Vincristine, vinblastine, bleomycin, idarubicin, epirubici Amsacrine, azacitidine, dideoxyinosine, dihydro -5-azacytidine, ethanidazole, ethiofos , Methotrexate, misonidazole, porphyromycin, pyrazolo acrizi Neck (pyrazoloacridinek), terephthalamidine, taxotere and other Taxane derivatives, topotecan, trimetrexate, N- Formyl-met-leu-phe-lys, arginine bradykinin, poly-L-lysine, other compounds Chemotactic triggers, biological response modifiers, cytokines, interferons, phosphorus Focaines and other agents useful in treating tumors or neoplastic diseases. the above Are used alone or in combination. In addition, in particular, treating infections Such active substances and methods include, but are not limited to: Not: gentamicin, amikacin, tobramycin, and other amines, salt Group, basic peptide, basic polypeptide, hydrophobic antibiotic, or amphoteric antibiotic Quality or bacterial disease, fungal disease, mycobacterial disease, viral disease Or an agent for treating other microbial or microbiological disorders.   The invention, in certain embodiments, can be described as a drug carrier composition, It has a sulfur content of up to 9% (w / w) and a selective oligosaccharide excess Including drug in combination with essentially purified dermatan sulfate with sulfation Wherein the composition has a non-embolic size of less than about 500 nm. Specific implementation In some embodiments, the drug can be a chelating agent. In certain embodiments, the composition The object has a size of less than about 250 nm. The drug carrier composition may also comprise a disease-inducing endothelium. By binding to the endothelium, the bound drug carrier composition within 10 minutes to less than 15 minutes If fully or partially covered, and up to 9% (w / w) sulfur content And having selective oligosaccharide oversulfation Dermatan sulfate purified to Ido-GalNAc4SO_ThreeAnd, furthermore, IdoA 2SO_Three-GalNAc4SO_ThreeAnd IdoAGalNAc4,6SO_ThreeCan be further defined when containing .   The drug carrier compositions of the present invention also, in certain embodiments, are nanoparticles. Can be defined as And this drug is preferably defined by a tumor treatment drug Can be The tumor treatment drug is preferably adriamycin, doxorubicin, d. From the group consisting of pirubicin, daunorubicin, idarubicin, or salts thereof Doxorubicin is most preferred. Alternatively, the tumor treatment drug Mycin, taxol, taxotere, vinblastine and vincristine, Amsacrine, azacitidine, dideoxyinosine, dihydro-5-azacitidine , Etanidazole, etiophos, methotrexate, misonidazole, porphy Lomycin, pyrazolo acridinect, terephthalamidine, taxotere, topo Consists of tecan, trimetrexate and carboplatin, or their salts It can be selected from a group.   A particular embodiment of the present invention is a drug carrier composition comprising doxorubicin, Selected from the group consisting of epirubicin, daunorubicin, and idarubicin The drug has a sulfur content of up to 9% (w / w) and is selectively oligosaccharide Contains in combination with essentially purified dermatan sulfate with oversulfation of Wherein the composition is less than about 500 nm, preferably less than 250 nm, most preferably It has a non-embolic size of less than 25 nm.   Embodiments of the present invention also include a drug carrier composition, wherein the drug comprises: An anti-infective (antiviral, antimicrobial, antifungal, or antituberculous) drug, Gentamicin, tobramycin or amikacin is preferred. Or this The drug is a biological response modifier (modifies an endogenous biological response) and is Alternatively, a biologically active peptide or polypeptide, and more preferably , Leukocyte chemotactic trigger, bradykinin, or poly-L-lysine. White blood The chemotactic chemoattractant is preferably N-formyl-met-leu-phe-lys (SEQ ID NO: 1). ).   The drug carrier composition of the present invention may be used in a pharmaceutical composition suitable for intravascular or other parenteral injection. Can be defined as being in a solution that is acceptable to the It can be formed by on-pair charge interactions, amphoteric or hydrophobic interactions.   In certain embodiments, the present invention relates to the treatment of tumors responsive to tumor therapeutics. And treating the animal by a method for preparing a drug carrier composition. (The composition has a tumor content of up to 9% (w / w) sulfur content, Purified derma with selective and selective oligosaccharide oversulfation Contained in combination with tansulfuric acid, where the carrier is non-embolic, 500 nm or less The above-mentioned drug carrier composition in a pharmaceutically acceptable carrier. Containing the drug carrier composition in a pharmaceutically acceptable carrier. Administering to an object. In preferred drug carrier compositions and methods The drug is in a controlled release form, and preferably A) Binding of a sample of the composition to the disease-induced endothelium results in endothelial induction, within 10-15 minutes In addition, it is understood that the bound sample is entirely or partially covered.   In embodiments involving treatment of various diseases, the drug carrier composition may be selected from Can be administered by isolated arterial perfusion, and can be administered in the proximal target organ, tissue or tissue lesion. Highly efficient uptake is obtained. Or, it can be administered intravenously, and Semi-selective, moderately efficient uptake of tissue lesions located in the body obtain.   The present invention is also a method of treating an animal for a tumor responsive to a tumor therapeutic. This method has a sulfur content of up to 9% (w / w), and Combined with essentially purified dermatan sulfate with over-sulfation of kalides To prepare a drug carrier composition containing a tumor therapeutic drug (here, The carrier has a non-embolic size of 500 nm or less); Containing in a pharmaceutically acceptable carrier; and a pharmaceutically acceptable carrier. Administering to the animal the drug carrier composition in the rear, wherein the tumor Therapeutic drugs are adriamycin, doxorubicin, epirubicin, daunorubicin , Idarubicin, bleomycin, taxol, taxotere, vinblastine And vincristine or a salt thereof.   Alternatively, the present invention provides a method of treating an animal for a tumor responsive to a tumor therapeutic. This method has a sulfur content of up to 9% (w / w) and Essentially purified dermatan with alternative oligosaccharide oversulfation A process for preparing a drug carrier composition containing a drug for treating tumor in combination with sulfuric acid. (Where the carrier has a non-embolic size of 500 nm or less); Containing the carrier composition in a pharmaceutically acceptable carrier; Administering to the animal a drug carrier composition in a pharmaceutically acceptable carrier. Here, the above-mentioned drug for treating tumor is amsacrine, azacitidine, dideoxy-inosi , Dihydro-5-azacytidine, etanidazole, etiophos, methotrexe G, misonidazole, porphyromycin, pyrazolo acrizinec, terephthal Amidine, taxotere, topotecan, trimetrexate, and carbopura Selected from the group consisting of chin or salts thereof.   In the specific treatment method of the present invention, the sample of the drug carrier composition is used to induce disease. Binding to the endothelium results in induction of the endothelium and the bound sample is totally or within 10-15 minutes. Alternatively, the drug carrier composition may be covered or partially covered by selected arterial perfusion. Highly efficient uptake in proximal target organs, tissues or tissue foci Is obtained. Or a group that is administered intravenously and is located at widely distributed systemic sites Obtain semi-selective, moderately efficient uptake in woven lesions.   In another embodiment, the present invention is a method of treating a vascular disease, comprising: Administering to the subject a therapeutically effective amount of a light drug, preferably the drug Contains metal ions.   Administration of the compositions of the present invention includes any mode of administration, including the targeted tumor of the therapeutic agent, Produces contact with the site of a disease or infection. It can be in a vein, artery, or cistern , Intraperitoneal, oral, or other modes of administration.   The compositions or formulations of the present invention may be in a pharmaceutically acceptable carrier or diluent. Or in any other pharmaceutically acceptable form. You. Such pharmaceutically acceptable formulations generally comprise a composition (eg, a pharmacology). Of doxorubicin (essentially purified dermatan sulfate) in typical preparations Containing amount.   The phrase “pharmaceutically or pharmacologically acceptable” when administered to a human, Presence and set of molecules that do not cause the reverse, allergic or other untoward reaction Say an adult. As used herein, a "pharmaceutically acceptable carrier" is Any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, It includes isotonic agents and absorption delaying agents. This for pharmaceutically active substances The use of suitable vehicles and agents is well known in the art. Any conventional media and drugs Unless the agent is incompatible with the active ingredient, its use in therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.   Accordingly, compositions of the present invention may be administered parenterally (eg, intravenously, subcutaneously, or intramuscularly). For oral administration (where the composition is a tablet, caplet, Or other solid). And the composition also comprises a target site. Depending on the location, time release capsules, and any other pharmaceutically currently used It can be formulated in a form (cream, lotion, gargle, inhalant, etc.).BRIEF DESCRIPTION OF THE FIGURES   The following drawings and figures illustrate preferred embodiments of the present invention and MRI contrast enhancement. Are presented to illustrate their use. These examples are purely illustrative And does not limit the full scope of the invention in any way.   Figure 1A shows control of diethylenetriaminetetraacetate (DTPA) substance 3 is an infrared spectrum (see Example 3).   FIG. 1B is an infrared spectrum of a control for L-lysine.HCl material (Example 3).   FIG. 1C shows a physical mixture of these DTPA and L-lysine.HCl substances (two substances Is the infrared spectrum of the control (without any chemical covalent bonds) (See Example 3).   FIG. 1D shows 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) bond. FIG. 9 is an experimental infrared spectrum of L-lysine covalently bound to DTPA in some cases (implementation See Example 3). Characteristic peaks in the 1250-1700 wavenumber range, indicating covalent bond formation Note the changes in height (height, width, and lack of division).   In the following figure (2-13), the dermatan sulfate carrier selectively Having an optimized oligosaccharide sequence but not undergoing total hypersulfation In a new special class of matane sulfate (SO_Three ^-/ COO^-Ratio = 1: 1, and sulfur Content = 6.3% by weight; Opocrin S.P. P. A., Corlo Di Formigine, Italy435 Ta Ip Supplied as ").   2A, 2B, 3A, 3B, 4A, 4B, 4C, 4D and 8A, 8B and FIG. 8C shows a ferrioxamine: dermatan sulfate selective magnetic contrast agent (Example) Before (Pre) and after (Post) intravenous (i.v.) injection (prepared in 2 and 5) T1-weighted MRI images performed at 1.0 Tesla and 1.5 Tesla (TR / TE = 800 / 45, 550/23 and 600/45) (tumor diameters at 1.0 cm and 1.5 cm at the time of imaging) Fisher 344 female with synchronous mammaryoma pre-inoculated into the liver as is between Rats were injected i.v. with ferrioxamine at a dose of 0.155 mmol / Kg).   FIG. 2A. Precontrast image of the liver (tumor is inconspicuous).   FIG. 2B. Ferrioxamine, a selective paramagnetic contrast agent: dermatan sulfate (0.15 5 minutes after intravenous injection (MPI) of 5 mmol / Kg) (MPI). Marked control enhancement of tumor in lobe, clear tumor border to surrounding liver, And isolated fraction, darker central area of tumor necrosis--tumor perfusion and organ Government can be spatially resolved and evaluated within various very small anatomical sub-regions. You.   FIG. 3A. Pre-contrast image of liver (tumor is present but not prominent) )   FIG. 3B. Ferrioxamine active only (including any dermatan sulfate carrier) Liver image at 7 MPI. Acute contrast enhancement is minimal Note that is or does not exist. This is clearly seen in FIG. 2B. Tumor enhancement, which is significantly different from dermatan sulfate carrier. The binding of the ferrioxamine active substance is associated with tumor site localization and ferrioxamine This is a requirement for tumor uptake of the active substance.   FIG. 4A. T1 pre-contrast image of liver (TR / TE = 800/45) (breast cancer present But not noticeable).   FIG. 4B. Ferrioxamine: dermatan sulfate selective MRI contrast agent at 21 MPI Liver image. Note major tumor mass and significant enhancement of distinct tumor boundaries . Small metastases (2 mm) in the left lobule of the liver, increased brightness thing. This transition is completely de-visualized in the pre-contrast T1 image.   FIG. 4C. Ferrioxamine: dermatan sulfate selective MRI contrast agent at 30MPI Liver image. Note the persistent enhancement of major tumors and metastases.   FIG. 4D. Ferrioxamine: dermatan sulfate selective MRI contrast agent at 42 MPI Liver image. Note: extended post-contra with highly persistent sensitivity Continuous strong enhancement of major tumors and metastases at strike intervals, and both measures With a continuous depiction of the tumor border in (selectivity), and further within 2 mm liver metastases Depiction of a very small non-reflux region at the center.   FIG. 4A, FIG. 4B, FIG. 4C, and FIG. Problem area (ROI) analysis of MRI image intensity from object analogs. Upper line is tumor ROI; lower line Is liver ROI; time point is pre-contrast; and ferrioxamine: dermatan sulfate The acid selective MRI contrast agents are 12, 27, 44, 64 MPI. The ratio of tumor intensity to liver Note that below: (a) 11: 1 at peak time of 12 MPI; (b) 27-64 MPI 3.2: 1 as an average over-furthermore, both (a) and (b) Always shows good selectivity. Kinetics reported against Gd: DTPA (this is Has a t1 / 2 at a site very fast, about 12-20 minutes (images not shown) ) Unlike, the intensity decays very slowly over time.   FIG. Syngeneic thoracic gland excised from the liver inoculation site of a female Fisher344 rat Histological staining of formalin-fixed sections of bean (heated ferroferricyanian) Reaction): ferrioxamine: dermatan sulfate selective MRI contrast agent external tumor Rim7 ~ 10MPI. (a) Strong positive on or in tumor endothelium, (b) Strong positive in subendothelium (C) moderate positives in the extracellular matrix of the tumor, and (d) Note the selective staining for moderately positive iron ferrioxamine.   FIG. 7A. Except for tissue sections, the same tumor, staining, conditions, and post-con Trust time, central tumor, ferrioxamine: dermatan sulfate selective MRI imaging Obtained from 7-10 MPI of the agent. Prominent staining positives are present at all sites in FIG. You.   FIG. 7B. Similar to FIG. 7A, various animals with the same type and site of breast tumor 6 and 7A except for the same ferrioxamine dose as in FIG. Only 7-10 MPI after i.v. for Samin active substance. Note the complete lack of positive staining. This It is the MRI imaging of the intact drug (active substance that binds to the carrier) versus the active substance Directly related to the results of MRI imaging of quality only (free active substance) (FIG. 2A And FIG. 2B vs. 3).   FIG. 8A. T1 weight of lung field in rats with primary hepatic breast tumor ( TR / TE = 600/45) Image. Pulmonary metastases (2 mm to 3 mm nodules) Note that it is a trust.   FIG. 8B. Lung field of the same rat at 12 MPI. Selective, bright in lung metastases Significant improvement in sensitivity of tumor detection (significant) due to enhanced sensitivity. Also, at the tumor border Pay attention to sharpness.   FIG. 8C. Same lung field of 17 MPI-persistent enhancement and sustained tumor boundary Sharp sharpness. By comparison, the fast diffusion rate of Gd: DTPA Rapidly induces fuzzy boundaries during anagen, and also reduces the sensitivity of detecting lung metastases I do.   9A, 9B, 9C, 9D, 9E, 10A, 10B, 10C, and 10D And FIG. 10E shows gadolinium injected intravenously at a dose of 0.100 mmol / Kg Gd (III). C DTPA dimeglumine (FIGS. 10A, 10B, 10C, 10D, 10E), Prepared in Examples 2 and 5 injected at a dose of 0.155 mmol / Kg iron (III) Ferrioxamine manufactured: Intravenous (i.v.) of dermatan sulfate selective paramagnetic contrast agent 9) Before (pre) and after (post) the injection (FIGS. 9A, 9B, 9C, 9D, FIG. 9E) shows a T1 weighted MRI image (TR / TE = 250/8) performed at 4.7 Tesla. Prepared in advance so that the diameter of the tumor is between 1.0 cm and 2.5 cm at the time of imaging Inoculated isotype of another AT-1 into a damaged skin pouch [Harn et al. (1993)] Each of these drugs administered to Copenhagen rats with prostate cancer.   FIG. 9A. Ferrioxamine: Precon for dermatan sulfate selective contrast agent Trust image.   FIG. 9B. Ferriox in liquid form at a ferrioxamine concentration of 0.166 mmol / mL Samin: 7 MPI of dermatan sulfate. External rims fan-shaped from the tumor stem Note the strong enhancement of the vascular array.   FIG. 9C. Same as FIG. 9B, except that it is 20 MPI. Persistence of elements in FIG. 9B Beware of discontinuous enhancements.   Figure 9D. Same as FIG. 9C, except that it is 40 MPI. Sustained connection of external rim Pay attention to trust and depiction.   FIG. 9E. Same as FIG. 9D, except that it is 60 MPI. Start of contrast decay Pay attention to the ball.   FIG. 10A. Pre-contrast image of Gd: DTPA dimeglumine non-selective contrast agent image.   FIG. 10B. Gd: 7MPI of DTPA dimeglumine. The outer rim is a very early post Note that even at the control interval, it is not well described.   FIG. 10C. Same as FIG. 10B, except that it is 20 MPI. Some seen in the center With significant drug sequestration, significant initial overall contrast decay reduces the area of the tumor. Mostly not circulating (cystic) Note that, as is typically reported for Gd: DTPA).   FIG. 10D. Same as FIG. 10C, except that it is 40 MPI. Almost backing up Note that we are back at the round level.   FIG. 10E. Same as FIG. 10D, except that it is 60 MPI. Except for the central cystic area Therefore, there is no reduction in the residual construct.   FIGS. 11A, 11B, 11C, and 11D show a dose of 0.155 mmol / Kg of iron (III). Followed by an acute 90-minute myocardial infarction (ligation of the proximal left anterior descending coronary artery) followed by injection of contrast agent Before injection, injected into a German Shepherd dog with reperfusion for about 90 minutes, Example 2 Ferrioxamine prepared as in 5 and 5: dermatan sulfate selective 0.5 te before (pre) and after (post) the rapid intravenous (i.v.) injection of magnetic contrast agent T1 weighted MRI ECG-gated cardiovascular images (TR / TE  = 250/8).   FIG. 11A. Pre-contrast image.   FIG. 11B. 7 MPI. Ferrioxamine: Strong enhancement of infarction by dermatan sulfate And in particular delineate the boundaries of the infarct--the boundaries of the putative peripheral region. Central Darker area-note the putative irreversible central infarct area.   FIG. 11C. 20 MPI, showing sustained strong enhancement and area above.   FIG. 11D. 40 MPI. Central area clogging, noticeable overall contrast Same as FIG. 11C, except that there is no attenuation. Note: (1) Ferrio at 0.155mmol / Kg Injection of the oxamine alone gives no detectable enhancement (image not shown); (2) Infarct size and location is verified by double staining injection immediately after imaging. It is.   FIGS. 12A, 12B, 12C, and 12D show a colony with an AT-1 prostate tumor model. MRI 4.7 Tesla T1-weighted images of Penhagen rats (FIGS. 9A, 9B, 9C, 9D, 9E, 10A, 10B, 10C, 10D and 10E). Show. However, rats were given lyophilized (versus liquid) forms of ferrioxamine: derma The tansulfate selective contrast agent is injected intravenously. And this medicine, 0.415m just before administration mol / mL Fe (III) was reconstituted to a higher concentration and 0.155 mmol Fe / Kg ( It is administered in the usual dose of III).   FIG. 12A. Ferrioxamine: Plecon for dermatan sulfate selective contrast agent Trust image.   FIG. 12B. Lyophilized fermentation reconstituted with water to a Fe (III) concentration of 0.415 mmol / mL Lioxamine: 7 MPI of dermatan sulfate. Very strong growth of the entire external rim of the tumor Be very careful.   FIG. 12C. Same as FIG. 12B, except that it is 20 MPI. External rim sustainability, non Always be aware of strong enhancements and depictions.   FIG. 12D. Same as FIG. 12C, except that it is 40 MPI. Start to brighten again Note the very strong enhancement of the external rim persistence associated with the central tumor. Also, in 40 minutes Note that there is virtually no contrast decay in the image.   13A, 13B, 13C, and 13D show Copenha with AT-1 prostate tumor model. MRI 4.7 Tesla T1-weighted image of xen rat (FIGS. 12A, 12B, 12C, and FIG. 12D). However, in rats, Gd (III): DTPA-Lys: dermatan sulfate The acid-selective contrast agent was injected intravenously in a liquid form pre-concentrated to 0.415 mmol / mL Gd (III) ( i.v.) injected and given in the usual dose of 0.155 mmol Gd (III) per kg.   FIG. 13A. Gd (III): DTPA-Lys: Precon for dermatan sulfate selective contrast agent Trust image.   FIG. 13B. Gd (III): DTPA-Lys: dermatan sulfate selective imaging at 0.415 mmol / mL 7MPI of agent. Note the very strong enhancement of the entire outer rim and central tumor. This is because Gd is higher than that of ferrioxamine chelate (R1 = 1.5 to 1.8 [mmol.sec] -1). : DTPA chelate (R1 = 4.3 [mmol.sec] -1) is consistent with the higher paramagnetic capacity.   FIG. 13C. Same as FIG. 13B, except that it is 20 MPI. Sustainable, very strong Pay attention to the external rim which is strengthened in contrast. Central vascular array (different from cystic isolation Note also the stronger enhancement of (as was done).   FIG. 13D. Same as FIG. 13C, except that it is 40 MPI. Ferrioxamine: Dell Except for absolute enhancement, which remains brighter or brighter in all areas compared to In 40 minutes, the sustained build-up of the outer rim is just starting to decay Be careful.   14A, 14B, 14C and 14D show Copenha with the AT-1 prostate tumor model. T1-weighted images of MRI 4.7 Tesla of xenrat (FIGS. 13A, 13B, 13C, 13 D). However, rats received intravenous ferrioxamine-selective contrast media. Inject intravenously. And here the active agent is not shared with persulfated dermatan sulfate Bindingly bound, the drug is lyophilized, and immediately before administration, 0.332 mmol / mL Reconstituted with water to a concentration of Fe (III) and 0.155 mmol Fe (III) per kg Is administered at a dose of   FIG. 14A. Pre-contrast.   FIG. 14B. 7 MPI.   FIG. 14C. 20 MPI.   FIG. 14D. Proportional to ferrioxamine bound to natural dermatan sulfate (above) At all times, slightly greater enhancement of the tumor rim and better vascular array Note the correspondence with the larger resolution.   FIGS. 15A, 15B, 15C, and 15D show a copy of the AT-1 prostate tumor model. MRI 4.7 Tesla T1-weight image of Penhagen rat (FIGS. 13A, 13B, 13C, And FIG. 13D). However, ferrioxamine selective contrast agents (where The active substance binds non-covalently to oversulfated chondroitin sulfate) in the rat Inject intravenously. The drug is then lyophilized and immediately before administration with water at 0.332 mmol / mL F Normal dose of 0.155 mmol Fe (III) per kg reconstituted to a concentration of e (III) Is administered.   Figure 15A. Pre-contrast.   FIG. 15B. 7 MPI.   FIG. 15C. 20 MPI.   FIG. 15D. 40 MPI. Ferrioxamine bound to natural dermatan sulfate (above) Medium greater enhancement of tumor margin at 7 MPI and greater vascular array Note the large resolution as well as the slightly larger enhancement at two later times .   FIGS. 16A, 16B, 16C, and 16D show the results of a colony with the AT-1 prostate tumor model. MRI 4.7 Tesla T1-weight image of Penhagen rat (Figure 13A, Figure 13B, Figure 13C, Figure And 13D). However, ferrioxamine selective contrast agents (where The active substance binds non-coagulant non-coagulant GAG, heparan sulfate non-covalently to rats I have an injection. The drug was lyophilized and immediately before administration 0.332 mmol / m L Reconstituted with water to a concentration of Fe (III) and passed 0.155 mmol of Fe (III) per kg It is administered at the usual dose.   FIG. 16A. Pre-contrast.   FIG. 16B. 7 MPI.   FIG. 16C. 20 MPI.   FIG. 16D. 40 MPI. The differentiation edge achieved by essentially all other GAG carriers (di fferential rim) at all actual post-contrast times proportional to the enhancement Note the very homogenous enhancement of the outer margin and central tumor. This property is It may be useful in diagnostic and / or therapeutic applications.   FIG. 17A shows gadolinium diethylene triamine pentaacetate (Gd: DTPA) 12 is a control infrared (IR) spectrum of Example 21 (see Example 21).   FIG. 17B shows a control IR spectrum of N-methyl-1,3-propanediamine (MPD). (See Example 21).   FIG. 17C shows the mixture of the individual chemical components Gd: DTPA and MPD (1: 1 molar ratio). 2 is a control IR spectrum of the (dry and dried) solution.   FIG. 17D shows covalent attachment to DTPA (as described in Example 21) in a 1: 1 molar ratio. 3 is an experimental IR spectrum of an MPD. Significant at 1400 wavenumber (signature ) Changes in peak height and separation, and wider expansion in wave numbers 3300-3600 Note the change in band height and configuration, which indicates covalent bond formation.   FIG. 18A shows that the tumor diameter at the time of image formation is between 1.0 cm and 2.5 cm. , Female Fischer 344 female with syngeneic thoracic adenocarcinoma previously inoculated into the liver Shows a T2 weight MRI scout image (TR / RE = 2100/85) of the liver area of the rat At 1.0 Tesla, just before running the T-1 weight series image (shown below) Have been. This T2 image shows two tumor nodules (right posterior l iver)) and identify the approximate location of one tumor infiltrate (central liver area) All tumor growth was confirmed at necropsy by rough visual inspection. It is.   18B, FIG. 18C, FIG. 18D, FIG. 18E, and FIG. And the diameter of the tumor at the time of imaging is between 1.0 cm and 2.5 cm. Fisher 34 with syngeneic thoracic adenocarcinoma previously inoculated into the liver 4 Gd: MPD-DTPA: de injected into female rats at a dose of 0.155 mmol / Kg according to Example 25. Before (pre-contrast) and intravenous (i.v.) injection of lumatan sulfate selective contrast agent Run at 1.0 Tesla at various times post-injection and post-injection (post-contrast, MPI) 2 shows the obtained T-1 weight image (TR / TR = 800/45).   FIG. 18B. Liver pre-contrast image (tumor not prominent)   FIG. 18C. T1 liver image 7MPI, Gd: MPD-DTPA: dermatan sulfate selective contrast agent (0.155 mmol / Kg). The same position as shown by the T2 weight scout image (FIG. 18A) Significantly better resolution of the tumor margin and higher control at the tumor margin (With last gradient), two solid tumor nodules (right posterior lobe of liver) And very strong contrast enhancement of one irregular tumor infiltrate (central liver area) Show strength. Improved resolution of small size tumor nodules and central tumor infiltrates Be careful. Both of these are T2 imaging artifacts, ie, the actual tumor margin (ma rgin) additional rim (corona) of water outside of the (But not in preferred T1 mode) but not present in T1 mode. Due to   Figures 18D and 18E. 20MPI and 40MPI, Gd: MPD-DTPA: Dermatan sulfate selective T1 liver image with contrast agent (0.155 mmol / Kg). This is because two solid tumor nodules ( Continuous emergency of the right posterior lobe of the liver) and one irregular tumor infiltrate (central liver area) With significant contrast enhancement, with a continuous, highly fractionated tumor margin And with essentially no contrast decay.   FIG. 18F. T1 liver images at 20 MPI and 40 MPI. This is two solid tumors Contiguous nodes (right posterior hepatic lobe) and one irregular tumor infiltrate (central liver area) Shows a very pronounced contrast enhancement and a sharpness of the tumor margin at 40 MPI Very slight attenuation, tumor control in two solid tumor nodules (right posterior lobe of the liver) Very small decrease in last intensity (decrease), further tumor infiltrates (central liver area) Increased brightness, and very slight background in the liver, unrelated to surroundings This is accompanied by an increase in brightness.   FIGS. 19A, 19B, 19C, 19D, and 19E show previously prepared skin. Has syngeneic AT-1 prostate cancer inoculated in the sac [Hahn et al., (1993)] and has 1.0-2.5 4.7-Tesla T1-weight fraction of Copenhagen rats imaged at cm diameter An image is shown.   FIG. 19A. Because surface coils are used and whole body coils are not used, Gd: MPD-DTPA: Dermatan sulfate selective, showing only back fat and back muscles Pre-contrast image for contrast agent.   FIG. 19B. Post-contrast image, 7MPI i.v. Gd: MPD-DTPA: Dermatan sulfate Selective contrast agent, liquid form. Extremely strong enhancement of the whole tumor mass and the tumor and below Note the extremely strong gradient (right image) at the border between certain normal tissues That.   FIG. 19C. Post-contrast image, Gd at 20 MPI i.v .: MPD-DTPA: dermatan sulfate Selective contrast agent, liquid form. Extremely strong enhancement of the whole tumor mass and the tumor and below Note the extremely strong contrast gradient at the boundary between certain normal tissues That. Contrast is slightly reduced in the central tumor area, and as a result Here, the tumor neovascular array is very well visualized.   Figures 19D and 19E. Gd: MPD-DTPA: Dermatan sulfate selective contrast agent (liquid Morphology) after contrast (40MPI and 60MPI). Still very strong growth of tumor Strong, especially very strong contrast in the connection between the tumor and the underlying tissue Note that the slope is maintained. Center tumor and outer margin (left image, motion Contrast strength) (apart from the object) is clearly such a very strong Gd: MPD-DTPA: Local high concentration of dermatan sulfate [R1 = 7.8 (mmol.sec)^-1] These areas Moderately reduced due to the accumulation of progressive tumors in T2^*Effect, center and outside Begin to develop competitive darkness in the lateral tumor area (left image, see also Example 26) ). The base edge (right side of the image) is relatively T2^*Protected from dark artifacts, This is due to the faster back diffusion of the drug into the plasma at the base. Therefore, moderate Indicates a lower dose.   FIG. 20 shows the results of the above and Example 26 and FIG. And freshly frozen, cut and stained tumor tissue as in FIGS. 6 and 7. Specific histochemical staining of AT-1 prostate cancer (from Copenhagen rats) Low-enhanced Prussian blue metal ion staining). Gd (I II) Note the selective staining positive for metal ions: (a) almost all Very strong positive in tumor tissue (intracellular site); (b) Strong positive in tumor cell nucleus-- Many tumor cells but not all tumor cells (for example, on the left side of the image, Tumor cells under the cell marker “9” and immediately after the grid marker “10” (See tumor cells on the left); (c) Moderately positive neovascular endothelial cells (eg For example, just above the grid marker "8" at the top of the image-"railroad t rack) "and see just below the grid marker" 2 " ); And (d) subendothelial and extracellular matrix sites (= tumor cells and endothelial ribbons) Weak positive to negative in the space between). Low 60 minute staining of extracellular matrix Can produce either (a) or (b) or both of the following: (a) in 60 minutes A more diffuse distribution of metal ions (for 7 minutes in FIGS. 6 and 7A), ( Diffuse metal alloys that are more difficult to visualize (because of their smaller optically stained nuclei) ON; or (b) plasma back-diffusion of a portion of the initially localized metal. These tumors Metal ions in tumor endothelium, tumor matrix, tumor cell-specific and tumor cell nuclei Positive findings indicate selective localization of MRI and radionuclide diagnostics and radioactivity Nuclide therapeutics, and in fact, provide the basis for other types of actives.   Figure 21A. 5 mg / Kg doxorubicin: DS (substantially purified dermatan sulfate Combined doxorubicin, type 435, Opocrin) at a weight ratio of 60:40 (doxorubicin) Host rats (Copenhagen stain) 3 hours after intravenous injection with syn: dermatan sulfate) Of prostate cancer (2/3 outside of tumor about 4cm in diameter), cut from color, syngeneic) Frozen 8-micron-thick sections and fireflies using rhodamine-type filters Light microscopy was performed to induce direct fluorescence of the doxorubicin drug substance (Example 29). See also). Packaged in relatively dense sheets in tissue sections Note very bright, direct drug fluorescence in almost all tumor cells . This indicates a high tumor cell internalization of the doxorubicin drug substance. Endothelial cytoplasm And nucleus positive (endothelial and tumor cell specific doxorubicin: DS (but not Non-canonical doxorubicin--see below) Note also).   Figure 21B. Same as the tumor in FIG.21A, but located around the endothelial stalk and located Subregions with slower clusters of tumor cells Tumor section (horizontally oriented across the image). Very bright in almost every cell Staining and doxorubicin localization in nuclear sites and tumor cell cytoplasm Notice the bright fluorescence. Also note the strong fluorescence of endothelial cells and endothelial cell nuclei Please.   Figure 21C. Copenhagen rats of the same tumor type and size but different (5 mg / K 3 hours after intravenous injection of g of standard doxorubicin (Adriamycin PFS liquid) Section). Dense sheet of cells in upper right and loose in lower left Note the proud clusters--all of which have significantly lower fluorescence (tumor and intracellular). Drug levels), as well as large tumor microvessels (center of image) Show a general lack of fluorescence at and around the No determinable fluorescence. The latter finding is a key intracellular site for drug action and Strongly lower drug levels at target (ie cell nucleus and nuclear DNA) Imply.                         Description of the preferred embodiment   The present invention relates to a non-toxic, biodegradable small molecule comprising an endothelial binding substance, in particular dermatan sulfate. Molecules, particles or microspheres (less than about 0.2-100 micrometers (μm) Size) and micro-aggregates (1-200 nanometers (nm)). this These substances have the following sequential steps when particles are injected intravenously into test rodents: Induces: 1) endothelial bioadhesion; 2) fast (2 min) endothelial coating of this material (partially Or whole); 3) Enhanced intact drug carrier particles across microvessels Moved into (accelerated) tissue compartment; (mostly completed within 10-20 minutes of injection And 4) coated carrier microsphere formulation (target tissue in vivo) (Known to correlate with the controlled bioavailability of the drug in the lesion) Delayed release of the drug.   The examples presented herein demonstrate efficient non-magnetic, normal and diseased tissue And compositions of matter that act as pharmaceutical carriers for sexual drug localization. This Of the specific classes of dermatan sulfate described herein (positive in the body). Binds to complementary heparin and heparan sulfate present on normal endothelium) Includes microspheres or nanospheres.   This material performs multiple endothelial connections and induces fast endothelial coating, and Association of microvascular beds, or potentially semi-selective (as proposed) systematic In any of the lesions of the disease after intravenous administration, enhanced macromolecules, Previously recognized in vivo that extravasation of aggregates and microparticles occurs And, if not indicated, the present invention relates to micro-fabrication from any currently proposed material. Considering that it is limited by the prior art including carrier matrix formulations Absent.   An endothelial-coated carrier can be formulated and can be either dry or fluid. And suitable pharmaceutically acceptable stabilizers, penetrants, Coloring agents, flavoring agents and ingredients that make them suitable for intravascular and intracavitary injection Physical solutions can be added. The present invention relates to external body barriers (eg, gastrointestinal tract, oral cavity, nasal , Rectal, sac or vaginal mucosa; efficient first-stage transfer across skin, cornea or sclera) Vascular targeting of any future device (see below) to develop in transport Consider the most specific application for a period.   The present disclosure relates to a drug key comprising microencapsulated spheres having surface adhesion properties. Carriers in tissues by endothelial bioadhesion and by induced endothelial migration, It shows that it was selectively taken into the tissue stroma. The present application further addresses such keys. The drug controlled by the carrier carries the drug in a selected target tissue (eg, lungs). It shows that it is deposited exactly in proportion to the deposition of the agent carrier. Now, even more soluble Drug-carrier conjugates (as well as formal microencapsulated drugs) To provide comparable tissue uptake of the drug under conditions where only Is set up. Now, furthermore, identical and similar carriers are found in the lungs, gastrointestinal tract and sac. It is established that it is taken up by the interepithelial pathway. Finally, the same and the same -Like carriers provide selective focus levels in tumors and foci of pulmonary vein infection Experience is established.   The unique aspects of drug carrier technology established by this application are Carrier, especially when the drug does not cause emboli (less than 3-4 μm) By providing efficient tissue uptake and localization of the drug when controlled is there. The carrier preferably has an embolus of less than 500 nm, more preferably less than about 250 nm. It does not wake up. Another unique feature is that these carriers are a) water soluble Formulated from a biocompatible, biocompatible and biodegradable material and b) a hydrophobic cap Tissue stroma (and lesion gel) in a manner not possible for the rear (eg, liposomes) It is widespread throughout. Finally, the carrier of the main embodiment is charcoal. Initial site of cell uptake based on carb-carbohydrate binding (endothelial and epithelial cells) Vesicles), and these interact in such a way as to give rise to multivalent bonds. Interact with each other, which can be both carrier and drug controlled by the carrier. Induced active endothelial (or epithelial) envelopment and endothelium penetration Leads to transit (or transepithelial) transport. This preferably involves transcytosis ( transcytosis), a process that occurs across a single endothelial or integumental cell Or include endothelial (epithelial) migration overgrowth of the carrier leading to encapsulation You.   In the practice of the preferred embodiment of the present invention, the active blood of the drug-carrier couple To induce extravasation (or epithelial transport), cells (or adjacent matrices) Polyvalent bond) must occur. Here, such transportation For uncoated (uncontrolled) particles or drug-carrier conjugates Compared to the transport obtained, it is significantly enhanced; this enhancement is achieved by non-embolic particles. Cell (complex) and embolizing particles (complex) transport through cells Blood flow and / or hollow fluid flow, air flow, or intestinal flow (each of Under in vivo conditions (in blood vessels, bladder, lungs, intestines or other body cavities), endothelium / Only within 12 minutes of epithelial contact (typically less than 5 minutes).   Carriers carry multiple (at least two) molecules of a drug through a simple transport Of various hormones, proteins, peptides and two low molecular weight drugs Delivery of these molecules must be controlled to distinguish them from conjugates.   The preferred embodiment describes a surface coating of dermatan sulfate, but another Rear (and surface coatings and drug-complexing agents), eg, dermatan Sulfate fragment, heparin fragment, tridodecyl methyl ammonium chloride Loride heparin (hereinafter TDMAC heparin), and other glycosaminog Lican (GAG), and preferably has a sulfur content of up to 9% (w / w), A new class of dermas that selectively oversulfate oligosaccharides Tansulfate also serves for constitutive and induced binding to heparin cofactor II. An 8-12 unit fragment of dermatan sulfate can activate heparin cofactor II. To join this. Unlike natural heparin, dermatan sulfate also has 8-12 Fragment does not inhibit constitutive endothelial surface clotting factor (antithrombin III) . This is also true for shorter semi-synthetic fragments of heparin. You. Therefore, both dermatan sulfate and both heparin and dermatan sulfate Fragments have lower anticoagulant activity than native heparin does It is foreseen. (Where natural heparin is the drug microsphere and complex The minimal anticoagulant activity of native heparin when incorporated into As low as possible. )   Endothelial uptake is a new physical formulation, dermatan sulfate and doxorubi A macromolecular complex with syn (an antitumor drug) is described. In this application Doxorubicin is combined with (preferably) the sodium salt form of dermatan sulfate To promote ion pair binding or complexation, preferably doxorubicin HCl It is understood that it is provided as. Accordingly, in the drug carrier composition of the present disclosure, When describing the active agent, doxorubicin and doxorubicin HCl are interchangeable Used for Noh. Selective and efficient uptake of this drug-carrier complex , In the present application, following administration of the conjugated agent. Dermatan sulfur It is also shown that there is no endothelial damage due to acid-doxorubicin. This new Good results using dermatan sulfate and related kit (as device) Established the principle for re-formulation of existing drugs. Reformulation immediately prior to drug administration Alternatively, it can be performed on site by a hospital pharmacist. This new app Is a drug-limited tissue controlled by a non-embolizing carrier ( Lesions) can allow uptake:   a) To the lungs (high efficiency delivery) and systemic lesion sites (moderate efficiency delivery) By intravenous administration; or   b) Selection for liver, kidney, brain, pelvis, limbs and other body parts (high efficiency delivery) By selective arterial perfusion.   The present application discloses that secondary tissue penetration of these hydrophilic drug carriers Normal target tissue (interstitial, lymphoid, etc.) for sulfate coated microspheres And what happens in the epithelium). In the present application, further examples are given. For lipid microemulsions, liposomes and other hydrophobic carriers In contrast to this situation, this hydrophilic sphere is used for tumor stromal and spontaneous pneumonia lesion gels. Penetrates widely, spreads the outer edges of these lesions and internal necrosis Establish a general principle of reaching both cores. This is an improved disease Cell invasion, cell (microbial) access and uptake of drug carriers, and It provides a new principle for trapped (controlled) drugs. This is Enables improved drug access to tumor cells and microbes at remote sites Then it is foreseen.   The present invention describes new entrapment of the following substances: Post:   a) doxorubicin HCl; and   b) Other antineoplastic drugs which are amenable to coating with dermatan sulfate and its derivatives (Eg, taxol, vincristine, or peptide tumor drugs).   The present invention relates to the use of internal drug nanoparticles, nanoemulsions, or internal drug emulsions. Trapped and controlled inside the formulation and other for entrapment Release excipients for preparing sub-capsules, complexes or drugs Formulations using the same surfactant. Such surfactants include: Including:   a) preferably sodium deoxycholate;   b) Alternatively, cholesterol, required for the formulation of stable drug emulsions , TWEEN 80, zwitterionic surfactant, or other biocompatible nonionic surfactant Surfactants, polysulphated surfactants, or positively charged surfactants.   The present invention relates to a method wherein cancer (and drugs) is improved by the use of the described technology. Teach that it can potentially be treated (and localized). Indicated in this application Bioadhesive carriers are highly toxic drugs (specific antitumor drugs); Unstable drugs; of large molecular size or of competing receptors scattered throughout the body Agents that experience inappropriate biodistribution or poor tissue access due to their presence; and And anti-adhesive drugs (prevent cancer cell metastasis, protect atherosclerosis, For the delivery of leukocytes and platelets to the vascular endothelium) It is expected to be favorable.   The present invention further provides for stabilizing the matrix and controlling the release of the drug. The use of the method. These are thickeners (eg, polylactic acid and polylactic acid). Glycolic acid, polyamino acid, poly-L-lysine, polyethyleneimine, glycero Or polyglycerol or polyalcohol (using heating or chemical reaction, Or without use), polyethylene oxide, biodegradable poloxamer or Is poloxamine (pluronic or tetronic) (Tetronic)), poly-COOH compound (polycarbol, or Polyamine).   Further methods of microparticulate formulation (especially for large-scale manufacturing purposes) Indicated as comprising the following method: preferably one (and / or coaxial) ) Of matrices through sonication or air flow disruption micropores (single or perforated) Extrusion of the composite (and / or surface) components; Aerosolization using a spray drying device.   The present invention relates to phase emulsification, and simultaneously matrix, surface and / or entrapment materials Solvent used for crystallization (preferably, hexane; or ethanol Or methanol).   Further sterilization (and / or particle sizing) of the final (or previous) preparation Preferably, for thermostable drugs, heating at 120 ° C. for 10-20 minutes Sterilization; preferably, for thermolabile drugs, submit the complex and nanoparticles. Clon filtration; and irradiation of particles larger than 0.22 μm; or ultrasound Including processing.   Many innovative teachings of the present invention will be described with particular reference to preferred embodiments of the invention. Put on. Here, these innovative teachings provide optimal spatial and dynamic -Specific localization and localization of paramagnetic metal chelates according to the chemical profile Of in vivo T1 type MRI image contrast enhancement It is advantageously applied to certain situations. On the other hand, at the same time, the Enhances clearance and minimizes its toxicity. But this key implementation It should be understood that these are merely examples of the many advantageous uses of the innovative teachings herein. It is. For example, the various types of innovative compositions disclosed herein and The method comprises a radionuclide imaging agent, an X-ray contrast agent, an ultrasonic acoustic image enhancer, And site delivery of metal chelates and in situ chelation of endogenous internal metals Localization of a wide range of therapeutic agents based on Can be used alternatively to enhance Therapeutic agents and agents included herein Of particular interest for tumor therapy, cardiovascular infarction, restenosis, atherosclerosis Sclerosis, acute thrombosis, microvascular disease, inflammation, and some of the onset and progression As described herein, the novel teachings, compositions and uses described herein provide binding, adhesion, In any other tissue disease with vascular components that are prone to metastasis and / or transformation Useful active substances and indicators. Thus, a number of additional compositions and uses It is obvious to a person skilled in the art that the invention can be uniquely implemented by the present invention. below An example is described in a preferred embodiment of the invention, its use in MRI contrast enhancement. It is shown to illustrate the use. These examples are purely illustrative, and It is not intended that the scope of the invention be limited in any way.   The present invention specifically relates to the preparation of novel contrast agents for magnetic resonance imaging And describe its use. These new contrast agents are distinguished from metal atom complexes Consisting of paramagnetic metal chelates. Here, the chelate disclosed herein is Binds to glycosaminoglycans (GAGs). The binding of the metal complex to GAG is an electrostatic phase Interactions (ion pairing), hydrogen bonding, van der Waals interactions, covalent bonds Or any combination of these interactions, but is not limited to these. Not determined. After parenteral administration of these metal complex-glycosaminoglycan preparations, The technology described herein utilizes a biocompatible carrier molecule to associate an associated biological The active substance is delivered to the site of vascular injury.   The present invention provides a substantially improved MRI image and spectral enhancement composition and Provide a way. These allow tumors, cardiovascular disease, and vascular or endothelial Of MRI hardware system for detecting other diseases having different adhesive components Ability to greatly increase. These improvements are achieved in the present invention by: You. That is, the chelated paramagnetic metal ions are selectively introduced to the target tissue site. Water protons or other diffusible nuclides present in the site (these nuclides are Orientation with constant and gradient magnetic fields and radio frequency fields at appropriate resonance frequencies (Localized) of T1 type paramagnetic relaxation (susceptible to pulse reorientation). Induces modulation and therefore either image contrast enhancement or spectral enhancement In the form, a detectable modulation of the induced magnetic resonance signal occurs.   Prior disclosure (Ranney: US patent application Ser. No. 07 / 880,660, filed May 8, 1992) US Patent Application No. 07 / 803,595 (filed April 3, 1992) and US Patent Application No. 07 / 803,595 / 642,033 (filed January 1, 1991) relates to the binding of magnetic materials that require: Do: (a) Greater than the magnetic power of gadolinium (III), the strongest single metal ion Magnetic ability; (b) a metal ion in which the bridging ligand disclosed herein is stably chelated; Non-bridging ligands with a higher degree of chemical and physical instability than Complex of intraatomically bound polyatomic metal atoms stabilized by: and (c) paramagnetism The invention provides for the only monovalent cationic charge on a metal chelator active. "Superparamagnetic" activity for binding to anionic carriers for the indicated requirements Divalent cationic charge on the material. For MRI use, metal chelators also Suitable paramagnetic metal ions (eg, preferably iron (III) or gadolinium (II I)), but with respect to certain other diagnostic and therapeutic compositions and uses. Thus, the metal chelators disclosed herein contain a suitable metal ion. It is also understood that this may be avoided. Preferred MRI application of the present invention , A basic metal chelator and a metal having electrophilicity at the formulation pH Chelators are preferred. For example, ferrioxamine (Crumbliss , 1991), basic or amine derivatives of polyaminocarboxylate chelators , Diethylenetriaminepentaacetate (DTPA), and macrocyclic chelators, 1, 4,7,10-Tetraazacyclododecane-N, N ', N ", N"'-tetraacetate (DOTA) salt Basic or amine derivatives (Li et al., 1993; Brechbiel et al., 1986). In certain cases Specific and powerful binding to these and related active substances Since the site localization can be emphasized by the carrier, the intrinsic property of the paramagnetic metal ion (a Paramagnetic R1) in vitro is the optimal site-localized image contrast or spectrum It is not essential to obtain the effect of the enhancement. Therefore, the present invention relates to basic metals Chelate, ferrioxamine (this is due to the chelated Fe (III) ion About 1.6 to 1.8 [mmol.sec]^-1T1 image connec Disclose the trust effect. Alternatively, the basic metal chelate of Gd (III) Under (but not all) in vivo conditions, the following can be expected. That is, when chelated by DTPA, about 4.0 to 4.3 [mmol.sec]^-1That Having potentially greater relaxation due to the larger in vitro R1 of And potentially moderately high when chelated by DOTA (Gera ldes et al., 1985), and when Gd (III) chelates to a particular DTPA derivative, R1 is 7.5 [mmol.sec]^-1High enough to be above. This derivative is a preferred DTP One preferred embodiment of the group of A-amine and DTPA-basic derivatives is N-meth Includes chill-1,3-propanediamine-DTPA. These include (a) water diffusion and mitigation Accelerated over DTPA and (b) acidic saccharides (preferably glycosamido (Including noglycans). Alternatively, metal ions are preferred Preferably, divalent or trivalent cations (manganese, chromium and dysprosium) ); And, less preferably, copper, nickel, erbium, -Cations of ropium and holmium.   The preferred chelators of the present invention are directed to the strong paramagnetic metal ions disclosed above. At least 10¹⁴A chelator having a formation constant of Contains a thionic group. These chelators are preferably ferrioxamine, Or DOTA, DTPA, porphine, porphyrin, sapphyrin or texaf Includes basic or amine derivatives of pilin. These are preferably Fe (III) and Capable of chelating, and most preferably chelating with Gd (III), and is essentially Can bind to the acidic groups of the acidic carrier by ion pairing (electrostatic) means. For example Certain texaphyrins have extended macrocycles, which, in certain cases, are Gd (III )) (Sessler et al., '065; Sessler et al.,' 720; Sessler et al., ' 498, incorporated herein by reference). Texaphyrin and sapophili Are not illustrated in the present invention, but are derived from the above cited references and herein. The illustrated and exemplified Fe (III) chelators (5,10,15,20-tetrakis (1-methyl-4- From pyridyl) -21,23-porphine), the following is clear to the person skilled in the art: Linked texaphyrins and sapphyrins and their basic, cationic And amine derivatives, and the porphine derivatives and the porphine derivatives disclosed herein. Analogs and basic, cationic and amine derivatives of And in combination with the acidic carriers disclosed herein and disclosed herein. Can be used. In particular, (a) paramagnetic performance of metal chelates; (b) binding to acidic carriers. And (c) formulation compatibility; and (d) in vivo biocompatibility and Alance is mutually considered. Hydrophilic chelators and carriers are Always (but not always) is preferred. It is made of those typical suitable Agent properties (no aggregation), biodistribution properties (hydrophobic plasma and No overall binding to the alveolar constituents); and clearance and poisons This is because of the superiority of sex. Alternatively, the chelator may be hydroxamate, ferri Chrome, enterobactin, felimicobacter, ferriclysin, and These basic or amine derivatives may be included. All these derivatives are Is defined to be subsumed below the original chelator.   A preferred carrier is an anion defined at the pH used for the formulation. Monomers, oligomers, and polymers containing or containing cationic or cationic groups. Contains limmer-like substances. These are typically carboxylic acid groups, and more preferably Or more preferably contain a stronger acidic group of phosphate groups, and most preferably a sulfate group. Includes. Preferred carriers include, but are not limited to, bacterial or semi-synthetic Typical saccharides of origin, acid saccharides, oligosaccharides, polysaccharides, Glycosaminoglycans or sulfatoids, or these Quality derivatives, modifications or fragments. All of these defined here Are included under the names of the original substances and categories above. Therefore, the preferred key Carriers include: heparin, desulfated heparin, glycine-bound heparin, Heparin sulfate, dermatan sulfate, chondroitin sulfate, pentosan polysulfate, And sulfated stalose (including sucrose octasulfate), and it Any derivative, modification, or modified form thereof. Most preferred are those herein Described above. Typical MRI preparations and Somewhat preferred for use are sulfated cyclodextrins, dextra Includes carriers of sulfuric acid and hyaluronic acid. But none of these May be particularly suitable for certain specific diagnostic or therapeutic formulations and uses.   In all cases reported and tested, basicity to acidic carriers Non-covalent binding of amine chelators is more likely to occur due to the covalent bond loading. This leads to significantly higher active substance loads. For example, preferred basic chelators (Ferrioxamine and Gd (III) DTPA-lysine, and most preferably, N- Chill-1,3-propanediamine-DTPA (N-MPD-DTPA)>70% by weight acidic To a glycosaminoglycan carrier. Another covalently active substance-carry A conjugate may be suitable in certain cases and for MRI applications. Are shown.   Certain embodiments of the present invention that have been tested in vivo have the following meanings as exemplified herein. Including but not limited to the following preferred embodiments: (a) deferoxami (Deferoxamine), (b) ferrioxamine, (c) Gd (III): DTPA-lysine, (d) N-meth Chill-1,3-propanediamine-DTPA, and (e) most preferred as exemplified below. Preferably by non-covalent means, and also preferably by covalent means , Other basic metal chelates bound to acidic glycosaminoglycans. This acidic The glycosaminoglycan is preferably dermatan sulfate (essentially 9% (w / w) And a selective oligosaccharide oversulfation ( purified dermatan sulfate with oversulfation), heparan sulfate and heparin is there. These may, by definition, reduce or improve anticoagulant activity. Site binding, enhanced clearance or other desired formulation or in vivo properties. Any derivative thereof, including hypersulfation and modification performed to provide Includes modifications. However, especially at higher doses, representing MRI contrast agent administration. Preferred carrier substances in view of low toxicity and optimal safety margin are preferably Between 0.7: 1 and 1.8: 1, most preferably between 0.9: 1 and 1.5: 1, and representative Target Has a 1: 1 ratio of relatively low SO_Three ^-/ COO^-And preferably 9% (w / w) Less than, most preferably between 4% and 7% (w / w), and typically 6.3-6.4% (w / w). dermatan sulfate which additionally has a relatively low sulfur content of (w / w). And The most preferred carrier material under the high dose administration conditions used above is also selected. Only selected oligosaccharide sequences are oversulfated, but the entire molecule is A new special class that is not oversulfated (as described and defined above) Dermatan sulfate. Alternatives apparent to those skilled in the art from the disclosure herein. Preferred agents are the basic derivatives of arginine and histidine of DTPA and DOTA And various texaphyrins, sapphyrins, porphines, porphyrins Filin, such a derivative of EHPG, may also be derived, and by definition, MRI Most preferably for their application contains their Gd (III) and Fe (III) metal ions And preferably also, as described above, their separate paramagnetic metal ion chelate. Contains a sheet.   The carrier material most preferably used in the present invention is a new class of essence Dermatan sulfate. It contains uronic acid (L-iduronic acid) Enriched and the sequence of its major monosulfated disaccharide (Ido- GalNAc4SO_Three), And oligosaccharides having a higher degree of sulfation. Also characterized by the sequence of the This sequence is Array ((IdoA2SO_Three-GalNAc4SO_Three) And (IdoAGalNAc4,6SO_Three)) (Disaccharide Assessed by analysis, and¹H and¹³Uniquely correlated with C magnetic resonance spectrum To). This purified dermatan sulfate is enriched in heparin cofactor II activity (Preferably greater than 220 units / mg). However, factor Xa and Low antithrombin III activity and overall anticoagulant activity (USP anticoagulant assay) Preferably less than 10%, and most preferably 5% of standard heparin Less), SO_Three ^-/ COO^-Low ratio (preferably in the range 0.7: 1 to 1.8: 1, and most Preferably in the range of 0.9: 1 to 1.5: 1) and low sulfur content (preferably Less than 9%, and most preferably in the range of 4% to 7%), and preferably 10,000 And between 23,000 and most preferably between 13,000 and 19,000 Daltons -The lower end of this molecular weight range, after intravenous administration, is Are generally important for the carrier to be highly retained in the vascular compartment of normal organs. is there. And the upper limit of this molecular weight range is the effective binding and uptake of disease sites. While important in general, very rapid blood clearance by the renal pathway is still important. Occurs. This is due to the low background of blood imaging and early time after injection It is important to achieve low body retention quickly.   The dermatan sulfate of the preceding paragraph may optionally be prepared by the following method : (A) animal organs or tissues (bovine mucosa, porcine skin or lung, and specific Preferably for use in Ming, including bovine mucosa), and The protein content containing papain at pH 5-7 for as short a time as possible to remove (B) a large screen functionalized with quaternary ammonium groups Particles containing a styrene-divinylbenzene matrix of the order 0.3 and 1.3 mm Pass through a strong anion (basic) exchange resin having a size range; (c) between 0.7 and 2.0 Eluting the sulfated polysaccharide using a molar salt neutral salt solution of (d) Divalent or trivalent metals (including copper, iron and calcium, and preferably copper) ) Crystallizes dermatan sulfate as a low solubility salt; (e) Chelex 100 type ( Reconversion to sodium salt via cation exchange resin containing Bio-Rad 143-5852) A crack in a strongly basic anion exchange resin functionalized with a quaternary ammonium group; Chromatography shows that oversulfated oligosaccharide sequences (above) Selective enrichment. Here, this resin is typically 10 microns or less. Having particle size and 2-8% cross-linking; (f) eluate by reverse osmosis Concentrate; and (g) freeze-dry the resulting liquid to form a fine white powder . One example of the above dermatan species (which is intended to be in any way the scope of the present invention) (Not intended to be limiting) As described (Mascellani et al., WO 93/05074 ( 1993), incorporated herein by reference; Mascellani et al., (1994), by reference. Dermatan sulfate (sulfate) subspecies. It contains. One of the most preferred examples of this subspecies of dermatan sulfate is Opocrin S. P.A. (Via Pacinotti, 3 I-41040 Corlo Di Rormigine, Italy) And supplied Type 435 bovine mucosal dermatan sulfate (sulfate). this Chromatography of charge-suppressed molecular sieves by UV absorbance analysis Has a formal molecular weight of about 17,500 to 18,000 daltons, as determined by It has a sulfur content of about 6.2-6.6% (weight percent). One excess sulfated The saccharide sequence ((IdoA2SO_Three-GalNAc4SO_Three) And (IdoAGalNAc4,6SO_Three))including , These dermatan sulfates of specific oligosaccharide sequences with a high degree of sulfation This low sulfur content occurs despite the selective enrichment in the acid. Its wealth Activation correlates with high heparin cofactor II activity.   In the description of the above two paragraphs, (a) uronic acid (L-iduronic acid) content and And the enrichment of the 2,4-disulfated disaccharide sequence is (b) preferably between 10,000 and 2 3,000 daltons, and most preferably a molecular weight in the range of 13,000-19,000 daltons And (c) low overall sulfur content, typically in the range of 4.5-7% by weight. Corresponding low SO_Three ^-/ COO^-In combination with the ratio, the surprising and expected Correlates with advantages not to be: (a) for example, tumor endothelial cells, tumor extracellular matrix Rapid disease site binding, localization, uptake and deep penetration of tumor and tumor cells Potential at ex vivo; and (b) low induced platelet aggregation, anticoagulation and bleeding Side effects-these are more highly sulfated and / or longer chains (larger Characteristic molecular weight). This carrier is sulfated , Hypersulfated and polysulfated glycosaminoglycans, and Natural and synthetic sulphated, oversulphated and polysulphated polysaccharides Rides and sulfatoids-most particularly, these are 10% or more USP heparin tie having a c content and in the range of 15-145 USP units / mg or more It has anticoagulant activity.   Preferred dermatan sulfate (above) and most preferred new special dermatan Sulfate subspecies (essentially purified as prepared by the special process described above) Is used as a site-selective diagnostic substance or as a drug carrier substance In some cases, the older, older dermatan sulfate (i.e., (a) (B) not prepared from the above isolation and purification process; or ( c) those not prepared from such an optional process, and Provides comparable enrichment and selective sulfation of small oligosaccharide sequences ) Are clearly distinguished from everything. When used as described above, These preferred, essentially purified dermatan sulfates are also From dermatan sulfate, not only are they structurally different, but also , Heparan sulfate and heparinoids (which bind to normal endothelium and have varying degrees of Undergoes ex vivo metabolism and interferes with rapid and complete blood and body clearance Are also clearly distinguished [Dawes et al., (1989), herein. Incorporated as a reference in the book]. Further, those skilled in the art will appreciate the new special Derma Tansulfate used as a site-selective diagnostic substance or as a drug carrier substance From non-dermatan sulfate classes of glycosaminoglycans, even more It is clear that there is a clear distinction. Where the non-dermatan sulfate class Glycosaminoglycans include: (a) chondroitin Sulfates A and C-which partition the uronic (L-iduronic) saccharide of dermatan sulfate No. [Walton et al., US Pat. No. 4,489,065; Maeda et al. (1993), both of which are incorporated herein by reference. (B) heparin--this is uron (L-iduron). It does not partition acid structures, but has high anticoagulant activity and high normal endothelial binding [Crem ers et al., (1994); Kalishevskaya et al., (1988), both of which are incorporated herein by reference. (C) Hyaluronic acid--This is a non-sulfated glycosaminoglycan. (D) all polysulfated glycosaminoglycans and oversulfated sulfates Ids (e.g., bacterial polysulfates, including pentosan polysulfate)-all of these Characteristically, it has a sulfur content of 10% or more, Significant in vivo safety due to induced platelet aggregation and cell membrane fallopian tube aeration / lysis Or act as a cofactor for such cell lysis, and Normal somatic cells and tumor cells and other pathogenic cells / organisms (eg, chondro Produced by itine polysulfate, resulting in chondroitin polysulfate-induced membrane damage , Specifically described as a direct toxic cofactor "opening" in tumor cells (Landsberger (1984)). Therefore, the present invention And a new special dermatan, which by itself has significant direct cell damage or Does not cause membrane damage, but instead uses dermatan sulfate carrier itself Without (alone) alone, rapid (3-7 minutes) selective binding of diseased endothelium, rapid (10-5 min) endothelial cell transport, tumor uptake, deep matrix penetration, and attachment Induces tumor cell internalization of the deposited diagnostic or drug-active substance and intermediates (eg, Damage either the skin) or the final (eg, tumor) target cells.   This new special class of dermatan sulfate is high in L-iduronic (uronic) acid Content (compared to low or no content in chondroitin sulfates A and C) And relatively low apparent molecular weight (typically 25,000 daltons) Chondroitin sulfates A and C equal to or greater than the apparent molecular weight of By contrast, most typically less than 25,000 daltons) It is clearly distinguished from acid types A and C. New special with relatively low molecular weight Dermatan sulfate is a carrier material for bound or associated active substances When used, it has at least three surprising and unexpected advantages: (a Very rapid blood clearing of carriers and active substances, mainly by the renal route (Typically at blood t 1/2 for about 20-120 minutes, very slow as a function of increasing dose) (B) only in vivo metabolism from minima to disappearance--standard hepa Major Contras for Phosphorus, Heparan Sulfate and Chondroitin Sulfates A and C (Which results in extremely low residual in vivo deposition or retention of carrier material). And c) maximal and rapid vascular egress through the disease site Induced and “permeability-enhanced” endothelium (eg, the largest disease site and tumor Induced by vascular endothelial growth factor / vascular permeability factor (VEGF / VPF) ), Uptake of bound or associated active substances and internalization of tumor cells ).   This new class of dermatan sulfate is an anticoagulant (heparin-type) active substance And in the absence of bleeding side effects Despite this, this new special class of dermatan sulfate is surprising. And unexpected benefits (non-covalent or covalently linked amines, Or the ability of chemically basic chelators or metal chelates to be highly effective and effective Acting as a simple in vivo carrier), Through the non-elevated vascular endothelium and the `` elevated '' vascular endothelium Selectively at the site of disease (including tumors) and at the same time Very rapid clearance of the non-targeted fraction of Preferentially, in the manner of increasing doses that only increase very slowly, It was not previously recognized and was not obvious to those skilled in the art. This has the side effect Provides not only reduced and low in vivo storage, but also the following additional advantages: : (A) static for enhancing in vivo contrast with associated paramagnetic metal chelates Very low imaging background very early after injection of intravenous administration; and And (b) the ability to refer to doses that increase with acceptable safety. these The surprising and unexpected advantages of the low-sensitivity imaging device and detection method For the use of in vivo magnetic resonance imaging (MRI) paramagnetic enhancement. is there. Therefore, deposit sufficient local site concentration of paramagnetic drug (about 50-100 μM) High intravenous doses of paramagnetic metal chelates (typically 0.1-0.3 mmol / kg range) need to be injected. This is further attributed to the carrier material (new Benefits of using a special dermatan sulfate (binding the active substance to the carrier) (Preferably allowing non-covalent methods). Therefore, one of the carriers Most covalently linked active substance-polymer systems (anti- (Including systems), typically 7-12% (w / w), Preferably more than 70% (active substance to weight% of [active substance + carrier]) bound An amount of active substance may be possible. Therefore, the new special dermatan sulfate The properties of constructable non-covalent (and covalent) formulations are required for administration Dermatan sulfate carrier is minimized, and effective The further surprising and unexpected nature of selective localization of doses at ex vivo Provide significant benefits.   The present invention provides a novel MRI contrast agent for enhancing solid tumors and cardiovascular infarction. Preparation and utilization are described. Contrast agents are strongly acidic substances (including polysulfated carriers). Consisting of a cationic or basic paramagnetic metal complex associated with Preferably, it contains glycosaminoglycans. Use any acidic glycosaminoglycan It is obvious to those skilled in the art that this can be done. Among the ion pair systems described below, Notable are the ferrioxamines with glycosaminoglycans, glycosamino DTPA-lysine with glycans, N-methyl-1,3-p with glycosaminoglycans Lopandiamine-DTPA, and most preferably, a new dermatan sulfate N-methyl-1,3-propanediamine-DTPA with a special subspecies.   In one particularly preferred embodiment, the essentially purified dermatan sulfate (435 types with an apparent molecular weight of 17,000-19,000, selectively oversulfated Gosaccharide and has heparin cofactor II activity at least at about 220 U / mg, Opocrin) non-covalently (or covalently) with the following oncology actives: Assemble, localize them to the site of the disease, and clear from the rest of the body Used to facilitate treatment: doxorubicin, adriamycin, taxo -Vincristine, Vinblastine, Bleomycin, Idarubicin (idaru bicin), epirubicin, amsacrine, azacitidine , Dideoxyinosine, dihydro-5-azacytidine, ethanidazo le), ethiofos, methotrexylate, misonidazole ), Porphyromycin, pyrazoloacridinec (pyrazoloacri) dinek), terephthalamidine, taxotere and other taxane derivatives , Topotecan, trimetrechelate, carboplatin , N-formyl-met-leu-phe-lys, arginine bradykinin, poly-L-lysine, etc. Chemoattractants, biological response modifiers, cytokines, interferons and Mfokine.   In another particularly preferred embodiment, the essentially purified dermatan sulfate (see 435 types with an apparent molecular weight of 17,000-19,000, selectively oversulfated oligos Opoc having heparin cofactor II activity at at least about 220 U / mg rin) is associated non-covalently (or covalently) with Yes: gentamicin, amikacin, tobramycin, and other amines , Basic, basic peptide, basic polypeptide, hydrophobic, or amphoteric Biomaterial or bacterial, fungal, mycobacterial, viral or other microscopic Biological or microbiological diseases.   Another diagnostic and therapeutic composition and use is in a set substantially similar to the compositions described above. It is envisioned that this can be done with an adult. For example, selectable metal ions are It can be chelated for metal-ion exchange at that site. Therefore, the present invention Formulations include manganese, aluminum, germanium, zinc, cobalt, and calcium. Metal, platinum, or other metal ions. Alternatively, irradiation For radionuclide therapy, such compositions may contain boron, cobalt, ruby, Indium, yttrium, technetium, ruthenium, rhenium, indium, Can it contain metal ions of ridium, thallium, samarium, or other Or may include. In particular, and in some preferred cases,⁵⁹Fe and⁶⁷ Ga [Hashimoto et al., 1983; Janoki et al., 1983] reported that radionuclei of non-radioactive metal ions Species forms include tumor, thrombus, and other nuclear medicine imaging forms for biomedical imaging purposes Can be replaced for production.   The above discussion is directed to the main aspects of the invention and its in vivo diagnostic and therapeutic applications. Presented to identify use in However, for those skilled in the art, Many additional and related compositions and methods of use are self-evident, And it is encompassed by the present invention.   The following examples are provided to illustrate preferred embodiments of the present invention. below The inventor of the present invention has made sure that the techniques disclosed in the examples function sufficiently to implement the present invention. On behalf of the technology discovered by them, and It should be recognized by those skilled in the art that the embodiments can be considered to constitute new embodiments. . However, one of ordinary skill in the art, in light of the present disclosure, will depart from the spirit and scope of the present invention. Many modifications may be made to the particular embodiment disclosed without departing from it, and It should be recognized that many changes can have similar or similar results It is.   In all of the following examples, unless otherwise specified, dermatan sulfate and heavenly Everything about natural dermatan sulphate contains only the selected oligosaccharide sequence. Has hypersulfation, but has hypersulfation of all molecules. New (as described and defined herein) Refers to lumatan sulfate, and especially Opocrin S.P.A., Via Pacinotti, 3, I-41040 Cor lo Di Formigine, a new special “435-type” model as supplied by Italy Refers to lumatan sulfate.                                 Example 1 Preparation of base-free deferoxamine and its use in the formulation of ferrioxamine   Base-free deferoxamine is used to minimize residual salt content present in the final formulation. It is used in certain examples to keep it low. This is a basic metal chelator And use deferoxamine. In these examples, deferoxamine is As previously reported (Ramirez et al., (1973), incorporated herein by reference). 2) KHCO_ThreeIs precipitated from the aqueous salt solution. Deferoxamine saturated solution Solution (320 mg / mL at 25 ° C.) in 12.5 mL of pharmaceutical grade water with 4.0 g of deferoxamine Prepared by dissolving the mesylate salt. Bring this solution to 4 ° C in a water bath Cool, and 2.5 mL of 2.0N KHCO_ThreeIs added. Stainless steel glass container Scratch with Le spatula and start precipitation. Collecting the precipitate by centrifugation; Wash repeatedly with ice-cold water and filter. Crude base-free deferoxamine , Purified by continuous recrystallization from hot methanol. Pure base-free obtained The deferoxamine is dried under a stream of nitrogen. Infrared of prepared deferoxamine The line spectrum is consistent with that referred to above.   Ferrioxamine is converted to a stoichiometric model of iron (III) and base-free deferoxamine. Formulated from base-free deferoxamine by the addition of iron (III) chloride, in the ratio of This produces chelated iron and minimizes residual mesylate and chloride ions. Reduce.                                 Example 2                   Preparation of ferrioxamine-iron (III) chelate   A batch quantity of the iron (III) chelate of deferoxamine is prepared as follows. 390 g of deferoxamine mesylate (methanesulfonate) (Ciba-Geigy Limited, B asel, Switzerland) is dissolved in pharmaceutical grade water. Or deferoki The chloride salt of samin may be used. Fe (O) OH (13.44% W / V Fe (O) OH particles, Noah Tec Highly purified iron (III) in the form of (hnologies Corporation, San Antonio, Texas) 372.9 g of the slurry are suspended in 1899 mL of water and, with constant stirring, deferro Add to oxamine. The resulting suspension is heated to 60 ° C for between 1 hour and 24 hours. And the pH is adjusted periodically between 6.5 and 7.9 by the addition of 0.10 N NaOH. Ferries Oxamine complex formation is evidenced by the development of a strong reddish-brown color in solution . Stoichiometric chelation of iron (III) with deferoxamine In-process (in-p) at 430 nm for chelated ferrioxamine standards rocess) Confirmed by using a UV visible light absorbance spectrometer. Cool batch solution to room temperature And centrifuged at 4500 rpm (about 2500 g) for 15 minutes to remove unreacted or aggregated Fe (O) OH Remove everything. Transfer this final batch volume as desired (generally a maximum of 2600 mL). Adjust to final volume). Remove any remaining traces of unreacted Fe (O) OH and remove Solution was also applied to a 0.22 μm Milli pore GV filter in a Class 100 laminar flow hood. Sterilize by passing the supernatant. Until the resulting batch is used further (See examples below), 4 in an autoclaved and capped glass container. Store at ° C. Determine the final concentration of ferrioxamine (DFe) by UV-visible absorbance spectroscopy at 430 nm Measure again by using the instrument. R1 = 1.6 (mmol.sec)^-1Is used for ICP-AA measurement of Fe (III) Based.                                 Example 3                       Preparation of basic amine chelator:         Diethylenetriaminepentaacetate-lysine (DTPA-Lys)   Dissolve 500mg DTPA in 20mL pharmaceutical grade water and heat to 60 ° C . 931 mg of L-lysine hydrochloride powder is added with constant stirring until dissolved. Ah Alternatively, N-ε-t-BOC-L-lysine can be converted to a carbodiimide intermediate at the lysine ε-amino group. (See below) and, if used, Methylformamide: Dissolve in water (50:50, w / v). Adjust the pH of this solution to 0.1N HCl Adjust to 4.75 by adding. 732.5 g of the water-soluble carbodiimide 1-d Dissolve tyl-3- (3-dimethylaminopropyl) carbodiimide HCl (EDC) in 2 mL of water And the pH is also adjusted as described above. While stirring this EDC solution constantly, The pH is periodically adjusted to 4.75 for at least 1 hour at 22 ° C and the DTPA lysine solution mixture (top And the reaction can be continued for a further 2 hours or more until complete. N-ε-t-B When OC-L-lysine is used (see above), the N-ε-t-BOC group is 1.0-2.0 During this step by acidifying with hydrochloric acid to a pH between and stirring for 30-60 minutes. Hydrolyzes. Readjust the pH to 4.75 as needed and rotate the reaction solution at 60 ° C Concentrate to 5 mL by Re-evaporation and DTPA-Lysine (DTPA-Lys) derivative Are precipitated by the addition of 3 parts by volume of ethanol. Note: Under these conditions, The ethanol: water ratio used maintains the solubility of all individual substrates (above) You. The resulting precipitate is collected by centrifugation at 2,500 xg, washed with ethanol, Centrifuge again and dry in a stream of dry nitrogen. Covalent attachment of lysine to DTPA Confirm by using an infrared (IR) spectrometer. The reaction product obtained is faint yellow Having.                                 Example 4               Preparation of gadolinium (III) metal chelate of DTPA-Lys                 : Gadolinium: DTPA-Lys [Gd (III): DTPA-Lys]   A gadolinium (III) chelate of DTPA-Lys, Gd (III): DTPA-Lys, is known Amount of DTPA-Lys is dissolved in water and until a stoichiometric amount of Gd (III) is added Add the required 0.1-1.0M gadolinium chloride stock solution To be prepared. The pH is adjusted to 7.0 by adding 1.0N NaOH. There Can add gadolinium oxide and stir the reaction mixture for 24 hours. Moth In the case of dolinium oxide, neutralization with 1.0 N NaOH is not required. Lys-DTP Each batch of A-conjugate is pre-titrated and standard xylenol orange titration ( Lyle et al. (1963)) was used to prepare the final chelation product for stoichiometric addition of Gd (III). And further investigation of gadolinium by using ICP atomic absorption spectroscopy Confirm by quantification. The obtained Gd (III): DTPA-Lys was converted to ethanol (1 volume part of water). (3 parts per volume) and the precipitate is recovered by centrifugation. You. The precipitate is washed again with ethanol and centrifuged (as above), Wash and centrifuge with Seton and dry the recovered precipitate in a stream of dry nitrogen. You. The resulting product has a similar faint yellow color as described in Example 3. to continue. R1 of aqueous product = 4.2 (mmol.sec)^-1Is based on ICP-AA measurement of Gd (III).                                 Example 5   Preparation of Ferrioxamine Ion-Pair Drugs Bound to Dermatan Sulfate Carrier       And ferrioxamine to depolymerized dermatan sulfate carrier   Ferrioxamine: dermatan sulfate ion-pairing agent is an aqueous solution of ferrioxamine Mix the liquid (see Example 2, above) and any of the following in the appropriate ratios Prepared by: (a) mode between about 5,000 and about 45,000 daltons MW dermatan sulfate (Opocrin, S.p.A., Modena, Italy, mode MW = 18,000 daltons 435 types of native bovine mucosa; and Scientific Protein Laboratories, Waunake, W insconsin, from porcine mucosa, mode MW = 19,600 daltons); or (b) about 2,000 daltons Depolymerized dermatan sulfate (Opocrin S.p.A. , Modena, Italy, depolymerized from bovine mucosa type 370, 435 type starting materials). De The range of the ratio of ferrioxamine to lumatan sulfate was reduced to 1:99 (wt%). Lioxamine: dermatan sulfate or depolymerized dermatan sulfate; and high 30:70 (wt%) )) Ferrioxamine: adjusted with dermatan sulfate or depolymerized dermatan sulfate To make. The pH of the mixture was adjusted to between 5.5 and 8 using 0.1-1.0 N NaOH, Is continued for 0.5-72 hours and the pH is re-adjusted between 5.5-8 (typically 7.5). adjust. Apply this ferrioxamine: dermatan mixture to a 0.22 μm filter To remove any residual insoluble iron oxides and hydroxides, Make the agent sterile. Sterile drug as a liquid at 4 ° C. or as a lyophilized powder ( (See below). For further processing, the liquid is Fill and autoclave at 120 ° C. for 15 minutes. There For further processing, fill the liquid into glass vials, freeze at -50 ° C, and By freeze-drying to produce a sterile freeze-dried powder. Lyophilized bath The vials were reconstituted by adding sterile water and mixing by hand for 1-5 minutes. Produces a reconstituted liquid of the desired concentration that is easy to inject. Ferrioxamine and de Measure the resulting concentration of lumatan sulfate and determine the vial volume using standard reverse phase HPLC. And polymer size exclusion HPLC method.   Multiple batches of ferrioxamine: dermatan sulfate drug were prepared. Typical The in vitro test results of the batch are as follows: ferrioxamine: de Rumatan sulfate ratio: 77.5: 22.5 (w / w); drug solubility, 550 mg / mL; water: octano Partition, 17,600 (± 2,750): 1; concentration of ferrioxamine, 0.166 mmol / mL; del Concentration of matan sulfate, 32mg / mL; molecular weight of dermatan sulfate, Opocrin 435 type, MN = 18,000 daltons; sulfuric acid / carboxylic acid ratio of dermatan sulfate, 1.0 ± 0.15; ferri Oxamine: dermatan purity, nominal ± 10%; pH, 6.5-7.9; viscosity, 3 .8-4.2 centipoise; permeability, 475-525 mOsm / Kg; R1, 1.5-1.8 [mmol.sec] -l; Extra large particles, small doses within USP guidelines for parenteral administration; anticoagulant activity, 4 .5 less than U.S.P.Units / mg (modified USP XXII assay); dermatan carrier At least 92% of the binding of ferrioxamine active against Dialysis for 3 hours against parts by volume, 500 dalton molecular weight separation).   In vitro stability of ferrioxamine: dermatan sulfate drug under enhanced conditions The gender indicates: (a) The liquid form is based on the above physicochemical and HPLC parameters Is stable for more than 6 months at 4 ° C, 22 ° C and 40 ° C; at 60 ° C Is slightly unstable in 2-6 months and at 80 ° C. in 1-3 days Very degraded. (b) liquid form with less than 3% degradation of ferrioxamine Can be autoclaved as described. (c) Freeze-dried forms include all, including oversized particles Is stable with respect to the above parameters (above); stable over several years of storage. Is shown.                                 Example 6           Preparation of ion-pair drug of ferrioxamine binding to heparin   Ferrioxamine: dermatan sulfate ion-pairing agent, ferrioxamine in water By mixing the liquid (as in Example 5 above) with the following in the appropriate ratio Prepare. (a) bovine lung heparin with a mode of MW between about 8,000 daltons; and (b) about 1 Porcine heparin with a mode of MW between 0,000 and about 20,000 daltons. Heparin Or lower the ratio of the ratio of ferrioxamine to heparin fragment, lower 1: 99 (wt / wt) ferrioxamine: heparin or heparin fragment; and high 3 Prepared between 0:70 (wt%) ferrioxamine: fragment. 0.1 ~ 1.0N N The pH of the mixture is adjusted between 5.5 and 8 with aOH and the mixture is allowed to run for 0.5 to 72 hours. Stir continuously and readjust the pH between 5.5 and 8. This ferrioxamine: The heparin mixture is passed through a 0.22 μm filter and the residual insoluble iron oxide-hydroxide And sterilize the liquid drug. Store sterile drug at 4 ° C. As indicated, further processing was performed by filling glass vials with sterile liquid and subsequent freezing. And lyophilization to make the drug into a sterile lyophilized powder. Lyophilized bath The vials were reconstituted by adding sterile water and mixing by hand for 1-5 minutes. Produces a reconstituted liquid of the desired concentration that is easy to inject. Ferrioxamine and f The resulting concentration of parin was measured and the vial volume was determined using standard reverse phase HPLC and Confirm by each of the polymer size exclusion HPLC methods.                                 Example 7           Preparation of non-anticoagulated heparin carrier by glycine derivatization   The anticoagulant activity of heparin was determined as reported (Danishefsky et al., (1971); Danishe fsky et al. (1972)) by derivatizing glycine residues and their carboxyl groups. And can be reduced to almost negligible activity. This non-anticoagulant Phosphorus (Nac-heparin) can be utilized as a modified glycosaminoglycan carrier. According to one method of glycine conjugation, weigh 0.75 g of heparin into a 100 mL beaker. Measure and dissolve in 25 mL of pharmaceutical grade water. Add 0.75g of glycine And adjust the pH of the resulting solution to 4.75 with 0.10N HCl. 0.75 g of 1-ethyl-3- ( (3-Dimethylaminopropyl) carbodiimide HCl (EDC) is weighed in separate vials. Weigh, solubilize by adding a minimum amount of water, and adjust the pH to 4.75 with 0.10 M HCl. Adjust to Aliquot an aliquot of EDC solution for at least 1 hour Add to the mixture of cans. After each EDC addition, adjust the pH to maintain at 4.75. You. After all the EDC has been added, stir constantly and adjust the pH periodically to allow the reaction to proceed. For a further 2 hours. The glycine-heparin conjugate (Gly-HEP) was then Precipitate by adding a volume of absolute ethanol. Precipitate 4500rpm (about 2500 × g) Collect by centrifugation for 15 minutes at Wash three times with ethanol.                                 Example 8 Glycosaminoglycans, modified and derivatized glycosaminoglycans (heparan sulfate Acid, non-coagulated heparin, excess sulfated dermatan sulfate, chondrotin sulfate, excess sulfate Binds to chondrotin sulfate and bacterial sulfatoids, pentosan polysulfate)                 Preparation of an ion pair drug for ferrioxamine   The ferrioxamine ion-pairing agent was converted to an aqueous ferrioxamine solution (see Example 5). Above) and the following glycosaminoglycans in an appropriate ratio Prepared using various glycosaminoglycan carriers: (a) MN = 8,500 Luton heparan sulfate; (b) non-coagulated heparin SPL, MN = 10,500 dalton ++; (c) MN = 19,000 daltons oversulfated dermatan sulfate; (d) MN = 23,400 daltons condo Rotin sulfate; (e) MN = 14,000 daltons oversulfated chondrotin sulfate and (f) MN = 2,000 dalton pentosan polysulfate. Glycosaminoglycans and sulfats The ratio of ferrioxamine to the id carrier is determined by the scaling factor ((mEq / mg carrier above) / (mEq sulfate / mg bovine lung heparin^★)), (Adjusted The effective payload ((ferrioxamine: carrier, 77.5: 22.5% (w / w)) Prepared as follows. Adjust the pH of the mixture to between 5.5 and 8 with 0.1-1.0 N NaOH; The mixture is continuously stirred for 0.5-72 hours and the pH is readjusted between 5.5-8. This Pass the ferrioxamine: heparin mixture through a 0.22 μm filter and remove the residual Remove all insoluble iron oxide-hydroxide and sterilize liquid drug. Sterile medicine Store the agent at 4 ° C. As indicated, further processing of sterile liquid into glass vials , Followed by freezing and lyophilization, and Make dry powder. Lyophilized vials are added to sterile water and manually Reconstitute by mixing to produce the desired concentration of reconstituted liquid that is easy to inject . The resulting concentration of ferrioxamine and heparin is measured and the vial The amounts are confirmed by standard reverse phase HPLC and size exclusion HPLC methods, respectively.   Although not prepared in the present application, the teachings of this example and the previous disclosure , 07 / 803,595, and 07 / 642,033 in combination with the ferrioxamine complex The body is purified with additional acidic saccharides (sucrose octasulfate and sulfated cyclode Glycans (including kistrin); additional glycosaminoglycans (keratan sulfate, hyaluro Acid); and additional sulfatoids (bacterial sulfatoids, dex (Including tran sulphate).^★ For bovine lung heparin, mEq SO_Three ^-/ g carrier = 4.4.                                 Example 9   Preparation of Gd (III): DTPA-Lys ion-pair drug binding to dermatan sulfate carrier   Gd (III): DTPA-Lys: dermatan sulfate ion-pairing drug at about 5,000 daltons and 45,000 MW of dermatan sulfate between 0 Daltons (as in Example 5 above) ), And in detail, dermatan sulfate with MN = 18,000 (Opocrin, S.p.A, Modena, Italy Prepared by mixing an aqueous solution of Gd (III): DTPA-Lys. 75: 25% (w / w) final solution of Gd (III): DTPA-Lys active against matane sulfate carrier Form a ratio. Prepare several stable drug variations of the resulting liquid I do. Here, the concentration of Gd (III): DTPA-Lys varies from 0.166 to 0.415 mmol / mL, The concentration of each dermatan sulfate varies between 35 and 87.5 mg / ml. Gd (III): DTPA-Ly T1 relativity of s (R1) = 4.2.                                 Example 10   Preparation of basic iron-porphine chelates; and ion-pair binding to heparin   5,10,15,20-tetrakis (1-methyl-4-), a soluble tetrabasic porphine Pyridyl) -21H-23-H porphine as a tetra-p-tosylate salt in 40 mg, 20 mL Reflux with 30 mg of iron (II) chloride in dimethylformamide for 2 hours. Iron complex Evolution evidence is observed in the development of red to dark green. Solvent is removed by evaporation The solid product is dissolved in water. Adjust the pH to 7.5, insolubilize excess iron (III), Thereafter, the iron porphine product is filtered. 2 mg / mL solution of iron-porphine complex And about 100% product yield confirmed by inductively coupled plasma atomic absorption . Comparable reactions in water yield about 70% yield.   This iron-porphine complex is converted to about 8 Kd, 1:20 to 20: 1 (iron-porphine: heparin). To bovine lung heparin dissolved in water. It has no precipitate A clear solution resulted. The binding of iron-porphine to heparin was 3.5 Kd and 12 Kd. Assessed by dialysis against water for 16 hours, using a bag with a molecular weight separation of Kd As is almost 100%. Alone iron porphine was almost completely dialyzed . UV-visible spectrophotometer measures 18: 1 (iron-porphine: heparin) molar ratio Shows the maximum binding yielding rate. The bovine lung heparin used was per mole (and Known to have about 18 effective strong acidic (sulfate) groups per heparin chain Therefore, these results indicate strong ionic interactions and bases on the sulfate groups of heparin. 2 shows stable (to dialysis) binding of a neutral tetraamine porphine complex.                                 Example 11         Preparation of basic triethylenetetraamine-iron chelate; and               Ion-pair binding to heparin and sucrose octasulfate.   1.0 g of triethylenetetraamine.2HCl (Syprine^TM) (Merck, West Point, PA) Was dissolved in water, and a 1: 1 molar ratio of iron chloride was added under acidic conditions (pH = 2), By obtaining a clear yellow solution, triethylenetetraamine and iron (III) To form a soluble complex of Adjust pH to 6.8 with 0.1N NaOH to form iron complex To give a red solution. This solution contains the iron-triethylenetetraamine complex. A fluffy red precipitate is formed which shows coagulation.   (a) To the aqueous dispersion of the obtained complex, bovine lung heparin was added, and the complex and heparin were added. The final ratio with phosphorus is between 95: 5 and 5:95 (weight). 65:35 ratio (complex: hepa At higher ratios of (phosphorus) and heparin, heparin completely solubilizes the complex . This apparent solubilization is due to the difference between triethylenetetraamine-iron and heparin. Shown are on-pair bonds.   (b) An aqueous dispersion of triethylenetetraamine-iron complex is added to sucrose octasulfate (SOS ) To bring the final ratio of complex to SOS between 95: 5 and 5:95 (weight). 65: 3 At a ratio of 5 (complex: SOS) and a higher ratio of SOS, SOS makes the dispersion very fine And shows an ion pair bond between the triethylenetetraamine-iron complex and SOS. Hepa The less complete clarification of this SOS ion-pair system compared to phosphorus (above) , A much higher density of SOS sulfate compared to heparin (this allows Hydrogen bonding between the subunits is substantially increased).   Although not directly illustrated, a family of C_xH_{x + y}N_xz(This is also iron (III) and cheap Polyamines which form a complex) are also the triethylenes of the present invention. It is clear that tetraamine-iron complexes and SOS can be used instead.   Preparation of covalent conjugate of deferoxaming glycosaminoglycan carrier   Substrates having an electrophilic amine group are described in [Danishefsky et al. (1971); (1972); Janoki et al. (1983); Axen (1974); Bartling et al. (1974); Lin et al. (1975)] As reported, acidic carriers, acidic saccharides, and acidic glycosamino A reagent that covalently binds to the nucleophilic carboxylate group of glycan. Can get. The coupling reagents described in these publications are carboxylic Activate the silate group. This mechanism is based on the carboxylate residue on the carrier. And the formation of an activated intermediate resulting from the reaction of the group with the coupling reagent. Middle The body is typically nucleophilically attacked by the amine function. This ensures stable sharing The formation of a covalent bond is typically via an amide bond between the active substance and the carrier. Occurs. Examples 12, 13, and 14 (below) describe ferrioxamine-heparin The synthesis of a covalent conjugate is described. Here, ferrioxamine has three distinct Covalently bound to heparin via a coupling reagent.                                 Example 12     By 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) bond               Preparation of covalent ferrioxamine-heparin conjugate   2.0 g of the aqueous ferrioxamine prepared in Example 1 is added with 0.10 M HCl Adjust to pH 4.75 with. Bovine lung heparin (Hepar-Kabi-Pharmacia, Franklin, OH) is dissolved in 5.0 mL of pharmaceutical grade water and with constant stirring Add to ferrioxamine. Readjust the pH of the resulting solution to 4.75 with 0.10 M HCl I do. 1-ethyl-3- (3-dimethylaminopropyl) which is a water-soluble carbodiimide 2 g of carbodiimide HCl (EDC) is weighed into a scintillation vial and a minimum amount of Solubilize with water and adjust the pH to 4.75 with 0.10 M HCl. Aliquot of EDC solution Pipetted to ferrioxamine-heparin mixture over 1 hour . After each addition of EDC, 0.10 M HCl is added to maintain the pH at 4.75. All of After adding EDC, the reaction is allowed to proceed for another 2 hours with constant stirring. Ferries The oxamine-heparin conjugate is prepared by adding 3 volumes of absolute ethanol. Allow to settle. The precipitate is centrifuged at 4500 rpm (about 2500 × g) for 15 minutes. And three times with 20 mL aliquots of ethanol and centrifugation Wash. The complex was redissolved in water and re-dissolved in 3 volumes of ethanol and centrifuged. Further purification by precipitation. Collect the final product and dry with nitrogen I do. The ferrioxamine derivative of heparin was converted to ferrioxamine at 430 nm. By UV-visible absorption spectroscopy and size exclusion HPLC chromatography Heparin Confirm by analysis.                                 Example 13     N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) bond           Of covalent ferrioxamine-heparin conjugate by HPLC   0.50 g of bovine lung heparin (Hepar-Kabi-Pharmacia, Franklin, OH)_TwoPurging Weigh into a 100 mL 3-neck round bottom flask equipped with an inlet and an outlet for the reservoir. Anhydrous dimethi 20 ml of formamide (DMF) are added with constant stirring and the resulting suspension Is warmed to 50 ° C. under a constant flow of nitrogen. 30 molar excess (about 463.7mg) of N-ethoxy Cycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) was added and the resulting The suspension is stirred at 50 ° C. for 3 hours. Activated EEDQ-activated heparin at 4500 rpm ( Collect by centrifugation at approximately 2500 × g) for 10 minutes. Pellets anhydrous DM Wash repeatedly with F, then three times with acetone. Activated intermediate with nitrogen stream Dry underneath.   Aliquot of ferrioxamine solution containing 766.3 mg of iron complex prepared in Example 1 Pipette into a 50 mL beaker and dilute to 25 mL with anhydrous DMF I do. In a separate 50 mL beaker, stir a known amount of EEDQ-activated heparin While suspending in 50 mL of anhydrous DMF. Add the DMF solution of ferrioxamine to EEDQ-F Pipette slowly into the palin suspension over 5 minutes. The resulting suspension Is continuously stirred at 40 ° C. for 3 hours. After cooling to room temperature, centrifuge and dry Wash 3 times with DMF, 3 times with acetone, and dry under nitrogen to obtain final product Collect things. Confirmation of formation of the conjugate is performed in the same manner as in Example 12.                                 Example 14             Covalent bonds via carbonyldiimidazole (CDI) bonds                   Preparation of ferrioxamine-heparin conjugate   The activating intermediate of bovine lung heparin (Hepar-Kabi-Pharmacia, Franklin, OH) was used for 3. Weigh 0 g of heparin into a 50 mL round bottom flask and add 25 mL of anhydrous dimethyl Prepared by adding formamide (DMF) with constant stirring. Carbo 608.1 mg (10 molar excess over heparin) of nil-diimidazole (CDI) Weigh into an vial and dissolve in 20 mL anhydrous DMF. DMF solution of CDI in DMF- Add to the heparin suspension and stir at 30 ° C. for 1 hour. After centrifugation, Wash repeatedly to remove unreacted CDI and residual DMF and dry under nitrogen. The CDI-activated heparin is recovered by drying.   Deferoxamine-heparin conjugate was added to 1.0 g CDI-activated heparin in 50 mL pills. By weighing into a bottom flask and suspending it in 25 mL of anhydrous DMF Prepared. Add 250 mg of deferoxamine prepared in Example 1 to another round bottom flask Weigh in and dissolve in 20 mL of anhydrous DMF. Deferoxamine free base solution The solution is slowly added to the CDI-heparin suspension and stirred continuously at 75 ° C for 16 hours. You. Centrifuge at 4500rpm (about 2500xg) for 15 minutes and wash repeatedly with anhydrous DMF Deferro by washing repeatedly with acetone and drying under nitrogen The oxamine-heparin conjugate is recovered. The resulting product is dissolved in water and Is measured by UV-visible spectroscopy. Stoichiometric amount of aqueous FeCl_ThreeAdd And the resulting solution is gradually adjusted to pH 6.5 and stirred for 2 hours. to this A deep reddish brown product is obtained. The ferrioxamine-heparin conjugate was added to 150 Dialysis against a volume of water through a 2,000 MW cut-off bag Separated from any remaining substrate and intermediates. Collect the retentate and rotary Concentrate by evaporation. Confirmation of derivatization is performed in the same manner as in Examples 12 and 13. .                                 Example 15       Covalent heparin-diethylenetriaminepentaacetate conjugate                           Preparation of (DTPA-heparin)   The DTPA-functionalized carrier was converted to diethylenetriaminepentaacetic acid dianhydride (cDTPAA; Ca lbiochem-Bhering Corp.) and molecules containing nucleophilic functional groups Prepared by 1.5 g of bovine lung heparin (Hepar-Kabi-Pharmacia, Franklin, OH) Dissolve in 5.0 mL of 0.5 M HEPES buffer and adjust the pH to 7.0 with 0.10 M NaOH. 4.5 g cDTPAA (about 100 molar excess over heparin) are weighed and 20 equivalents (22 Aliquots of 5 mg). Aliquots of cDTPAA are added until all cDTPAA has been added. And add to the heparin solution every 3-5 minutes. Adjust the pH of the solution during the addition of cDTPAA. Monitor continuously and maintain pH 7.0 with 0.10 M NaOH. cDTPAA last After adding an aliquot, the solution is stirred for a further 30 minutes. DTPA-heparin solution 1 Dialysis through a 1000 MW bag for 50 volumes to remove unbound DTPA Leave. The obtained conjugate was concentrated by evaporating with nitrogen at 37 ° C. And store at 4 ° C.                                 Example 16           Gadolinium (III) and DTPA-heparin covalent conjugate                           Preparation of iron (III) chelates   The DTPA-heparin conjugate of Example 15 was combined with the required amount of DTPA-heparin in 125 mL Pipetted into a Meyer flask and a 1.5-10 molar excess of paramagnetic gold. Gd Group Ion Oxide_TwoO_ThreeOr added as Fe (O) OH, and at 37 ° C. for 24 hours to 36 hours Stirring to obtain a solubilized metal oxide sufficient for full occupation of DTPA groups. Form of paramagnetic metal chelate of DTPA group with gadolinium (III) or Fe (III) It is further prepared in a state. The remaining metal oxide is centrifuged at 4500 rpm (about 2500 g) And the product is filtered using a Millipore 0.22 μm GV-type filter. Unreacted metal acid by filtration through Separation from chemical compounds. The chelated metal ion and heparin concentrations should be It is measured by inductively coupled plasma (ICP) and size exclusion HPLC. Gd (III) , Stoichiometric chelation and standard xylenol orange titration [Lyle et al. (1963) ].                                 Example 17         Ferrioxamine: dermatan sulfate, 435 type toxicity study   Acute intravenous toxicity studies using day 14 recovery and necropsy were performed in male and female rats. And in male and female dogs. Standard i.v. injection rate of 0.075 mmol / Kg / min In degrees, prominent signs are generally 5 to 12.5 times 0.155 mmol / Kg of the effective imaging dose. Occurs only after LD50 is much greater than 4.5 mmol / Kg, and tail-vein injection (tail-vein infusion). At this speed, several rats Can be injected at 10 mmol / Kg without undue effect. 0.080mmol Rats can die at an artificially accelerated i.v. injection rate of / Kg. And LD50 It is between 2.5 mmol / Kg and 3.0 mmol / Kg. At final necropsy, 2.2 mmol / Kg, 3.0 mm ol / Kg and 4.5 mmol / Kg after i.v. (N = 5 males and 6 females (per dose level)).   Pyramid acute i.v. toxicity studies were performed at 0.5 mmol / Kg, 1.2 in protocol studies. escalating doses of 0.01 mmol / Kg and 2.25 mmol / Kg and 0.012 mmol Performed on dogs at an infusion rate of / Kg / min. Complex of acute signs of hypotension (compl ex) can be obtained. It is minimal and reversible. Died, and 14 days Final necropsy of the eyes showed no abnormalities (n = 2 males and 2 females) . All received each of the three dose levels with a 72-hour rest interval . ).                                 Example 18               Ferrioxamine: dermatan sulfate selective contrast agent:        MRI imaging of lactating breast cancer in syngeneic Fisher 344 female rats;                   And its interaction with specific histochemical studies   2A, 2B, 3A, 3B, 4A, 4B, 4C, and 4D. As shown, Tl-weighted MRI images (TR / TE-800 / 45 and 550/23) , Ferrioxamine: dermatan sulfate, 435 type selective paramagnetic contrast agent (Example 5) at a dose of 0.155 mmol / Kg of ferrioxamine in Fisher 344 female rats (trocar Have a syngeneic breast cancer that has been inoculated into the liver with the liver. As a result, tumors during image formation Has a diameter between 1.0 cm and 2.5 cm. ) Before (pre) and intravenous (i.v.) injection Later (post) at 1.0 Tesla and 1.5 Tesla. Tumors have a standard Tl load Inconspicuous in heavy pre-contrast images. Ferrioxamine: dermatan sulfate After injection of the drug, tumors were rapidly and markedly enhanced (a) early (7 minutes) after injection. (FIG. 2A, FIG. 2B); (b) Very clear tumor border to surrounding liver (FIGS. 2A, 2, 4A, 4B, 4C, and 4D) and separately The darker central area of the necrotic tumor (FIGS. 2, 2B), And function are spatially decomposed into various very small anatomical sub-regions. (C) continuous, very long-lasting imaging time intervals. Sustained contras longer than 64 minutes after injection (MPI), with tumor boundaries defined in minutes Strike (FIGS. 4A, 4B, 4C, 4D, MRI images; FIG. 5, determination of region of interest (ROI)) Quantitative analysis). Microwave amplified iron staining (microwav) of newly dissected 7 MPI tumor The correlation of these MRI images with e. augmented iron stain Localization of the tumor site of the min-active substance, which binds (non-covalently) to the carrier And only in the free form (active substance alone) It does not occur when administered (FIGS. 3A, 3B). 8A and 8B As shown in FIG. 8C and in FIG. Rapidly and sharply enhanced at an early post-contrast interval within a 3 mm nodule; and Tumor enhancement at this lung site also results in very clear tumor boundaries for normal lungs. Long-term high sensitivity and retention of field lines. 8A, 8B and 8C. Gd: DTPA dimeglumine control at body organ sites It is much longer than typically reported for last enhancement.                                 Example 19     Ferrioxamine: dermatan sulfate selective contrast agent: Syngeneic Copenhagen           (Copenhagen) MRI imaging of prostate At-1 cancer in rats                         Comparison with Gd (III) DTPA   9A, 9B, 9C, 9D, 9E, 10A, 10B, 10C, As shown in FIGS. 10D and 10E, a Tl-weighted MRI image (TR / TE-250 / 8) Ferrioxamine prepared as in Examples 2 and 5 at 4.7 Tesla: Derma Before (pre) and intravenous (i.v.) injection of tansulfate, 435 type selective paramagnetic contrast agent And a post injection and an i.v. injection with a 0.155 mmol / Kg iron (III) dose (FIGS. 9A, 9B, 9C, 9D, 9E); gadolinium DTPA dimegumi Injection was performed at a Gd (III) dose of 0.100 mmol / Kg compared to 10B, FIG. 10C, FIG. 10D, FIG. 10E); Copenhagge with syngeneic AT-1 prostate cancer inoculated in a manufactured epidermal pouch Rats (Hahn et al.). As a result, the diameter of the tumor at the time of image formation was 1.0 cm and 2 It is between .5cm. Ferrioxamine: dermatan sulfate rapidly spreads tumor margins Strongly enhances and also fan out of the tumor stem, which carries most of the tumor vasculature It rapidly and strongly enhances the expanding vascular array. Sustained contra of these elements The strikes and depictions will still exist through the 40 minute dynamic point in time. By comparison For example, after Gd: DTPA dimeglumine, the outer edge is an early interval after contrast (7MPI) Not even enough. Significantly rapid loss of contrast was observed at 20 MPI. It occurs throughout the tumor and some drugs (such as those used for imaging Most of the center of the tumor (as typically reported for Gd: DTPA in combination) Isolated in non-perfused (cystic) areas. At 40 MPI, enhancement is essentially back Back to ground level, and at 60 MPI, except for the central cystic area No contrast remains.                                 Example 20       Ferrioxamine: dermatan sulfate causes acute myocardial infarction in dogs                           MRI contrast enhancement   As shown in FIG. 11A, FIG. 11B, FIG. 11C, and FIG. ECG gated cardiovascular imaging at 0.5 Tesla, ferrioxamine: Derma Before rapid intravenous (i.v.) injection of tansulfate, 435 type selective paramagnetic contrast agent (pre) And later (post). This is an iron (III) dose of 0.155 mmol / Kg, Germanche with intermyocardial infarction (ligation of the proximal left anterior descending coronary artery) Padd dogs are given an i.v. injection and then reperfused for approximately 90 minutes before injection of contrast agent . At 7 MPI, ferrioxamine: dermatan enhances infarct zone, especially infarct To distinguish the outer boundary of This outer boundary is a potential Represents the putative marginal band of infarct following recovery. The darker region in the center is the estimated irreversible Represents a large central infarction. Sustained strong enhancement and zoning are preserved through 40 MPI. Exist. Ferrioxami injected at 0.155 mmol / kg without carrier Shows no detectable enhancement. In these studies, infarct size and Position confirmed by double dye injection performed immediately after MRI imaging Is done.                                 Example 21     Ferrioxamine activity bound to various sulfated glycosaminoglycans               In vivo MRI tumor-imaging ability comparison of substances   Based on low anticoagulant activity, safety and projected site-localization potential Specific selectable glycosaminoglycan carriers and the resulting selective MRI imaging The specific selectable physical form of the agent is combined with the carrier medium of the associated ferrioxamine. The relative potency of in vivo tumor localization is compared. Subtleties in drug localization High spatial resolution and capacity to detect large quantitative differences, The AT-1 prostate cancer model of Example 19 is used.                               Table 2 (continued)   Carriers with a shorter chain length than glycosaminoglycans (ie, pentosan poly) Sulfuric acid) has a lower capacity (typically only 2/6 on the scale) And are found to remain at the tumor site at intervals of less than about 20 minutes. Place However, the GAGs shown in the table above have very large capacities and are significantly longer With the spacing of localization. Comparing these carriers, the carrier sulfate charge Increasing carrier capacity based on charge density, from slight to gentle slope There is a tendency of "slight-to-moderate". Lyo = lyophilized powder form SO_Three ^-= Sulfate (eg, Dermatan SO_Three ^-= Dermatan sulfate)^* Bovine mucosa, purified, 18,000 daltons^** Pig mucosa, 19,600 daltons                                 Example 22       Preparation of N-methyl-1,3-propanediamine derivative of DTPA (MPD-DTPA)                 And chelation with gadolinium (III)   Diethylenetriamine-pentaacetic anhydride (DTPA anhydride) solution was added to 180 ml of anhydrous Prepared by adding dimethylformamide (DMF) to a 250 ml round bottom flask I do. Attach the side arm addition funnel to the flask and add Put a magnetic stirrer into the lever flask. 5g while stirring DMF vigorously (14 mmol) of DTPA anhydride (Sigma Chemical Co.) in 0.5 g portions over one hour Added. Warm the resulting suspension to 60 ° C for 15 minutes or until the solution is clear I do. Remove flask from heat and place in ice bath until solution equals 4 ° C Put on.   MPD-DTPA derivative was combined with 15 ml of DMF and 1.46 ml (14 mmol) of N-methyl-1,3-propanediamine. Prepared by mixing with an amine (Sigma Chemical Co.) in an addition funnel. The MPD-DMP mixture in the side-arm addition funnel was stirred vigorously with cooled (4 ° C) DTPA anhydride. Add dropwise to the solution. A white precipitate is formed upon addition. Suspension overnight at room temperature And stir. The MPD-DTPA derivative was collected by centrifugation at 2500 g for 10 minutes, Wash repeatedly (5 x 300 ml).   In a concentrated solution, the product at this stage has a pH of 3.5 and for further purification a solution of pH 7.0 is required. Dissolve the MPD-DTPA derivative product in water and adjust the pH using 5N NaOH. Adjust to 7. The product is lyophilized for 16 hours and dried. Minimum amount of lyophilized material (40 Dissolve in warm (50 ° C) methanol for 15 minutes, cool to room temperature, and add 10 volumes Precipitate with acetone. The precipitate is collected by centrifugation at 2500 g for 10 minutes. This Substance was again dissolved in warm methanol for 15 minutes and precipitated with 10 volumes of acetone. And collected by centrifugation at 2500 × g. Repeat the precipitate with acetone Wash, dry under nitrogen, and store in a vacuum desiccator.   Infrared (IR) spectroscopy analysis of MPD-DTPA conjugate formation (see Figures 17A, 17B, and 17C) ) And HPLC chromatography. HPLC characterization Use a thione exchange column (Dionex IonPac CS14, 4 × 250 mm, 8 μm, carboxylic acid) Using 20 mM methanesulfonic acid in acetonitrile-water (99: 1), pH 1.8. Performed with UV detection at 220 nm using a mobile phase. This ensures good separation Chromatographically pure (> 99% purity) peaks: (a) DTPA (3.7 min); (b) N-methyl-1,1-propanediamine (MPD versus D required for detection The molar ratio of TPA is 20: 1. (8.4 minutes) due to low UV absorbance of MPD; (c) DTPA (also Is a mixture solution of hydrolyzed DTPA anhydride) and MPD (1: 1 molar ratio) (3.7 min ) (Only DTPA is detected; MPD is due to the very low extinction coefficient of MPD); and (D) Given for MPD-DTPA conjugate (1: 1 molar ratio) (15.6 min). (D )) Is> 93% according to HPLC absorbance at 220 nm.   The chelating ability of N-methyl-1,3-propanediamine-DTPA (MPD-DTPA) Lenol orange (5%, w / v) was used as a colorimetric indicator of endpoint and 1 M 0.1M GdCl in monium (pH 5.5) buffer_Three 5H_TwoTit small aliquots with O Is determined by Based on this titration, a stoichiometric amount of 1 M GdCl_Three 5H_TwoO Add to the batch volume of N-MPD-DTPA as below: bulk MPD-DTPA with minimal amount of water (approx. 300 mg / ml) and add 1M GdCl while stirring vigorously._Three 5H_TwoAdd O, and pH Is adjusted to <4.0-7.0 with 5N NaOH. The average chelating ability is about 22% by weight Yes, with slight variations due to the extremely hygroscopic nature of the dry chelators.                               Example 23       Preparation of gadolinium: MPD-DTPA: ion pair formulation of dermatan sulfate   Gadolinium (Gd): MPD-DTPA: dermatan sulfate ion-pair formulation, (new Gd: MPD-D of 10: 1 to 1:10 using special grade 435 type dermatan sulfate and Opocrin Prepared over a range of weight ratios of TPA to deltaman sulfate and, in particular, 60% Gd: MPD-DTPA to 40% deltaman sulfate (w / w), one of the new ratio ranges (Equivalent to a molar ratio of 43: 1). These ion pair preparations provide the desired amount of data. Rutaman sulfate was dissolved at a concentration of 400 mg / ml and Gd: MPD- prepared in Example 22. Prepared by stirring in DTPA. This enables laser light scattering analysis (Nico Completely hydrophilic, without molecular aggregates detectable by mp Instrument To A clear solution is obtained. Strong ion-pairing between GdMPD-DTPA and dermatan sulfate Is confirmed by dialysis (150 volumes) with a 500 MW cut-off bag for 3 hours. And the retained Gd is evaluated by ICP atomic absorption spectrometry (mass balance = 9 Five%). Gd: MPD-DTPA: Del prepared with 60: 40% (Gd: MPD-DTPA vs dermatan sulfate) For the matane sulphate formulation, a very strong ion-pair bond, 73% of GD in the bag Indicated by retention; prepared with same molar ratio of Gd: DTPA to dermatan sulfate Gd: DTPA: dermatan sulfate for GD in the bag Retention is very low at 23%.   Quantitation of dermatan sulfate was determined as previously described [Klein et al. (1982): Grant (1984), both of which are hereby incorporated by reference], the UC absorbance at 620 nm. (This is a very strong binding (substituting) cationic dye, Azure A , Which occur during the binding of   (A) Gd: MPD-DTPA alone and (b) Gd: MPD-DTPA: 60% of dermatan sulfate: 40 % (W / w) of the R1 titer (T1 relaxivity) of the ion pair formulation was determined by IBM PC20 Min Evaluated using an ispectrometer, and both are 7.8 mmol^-1Second^-1(IC Based on parallel measurement of Gd concentration by P atomic absorption). Gd chelate alone and Dell Equal R1 for the Gd chelate bound to the tansulphate is Shows that binding to matan sulfate does not interfere with water diffusion and paramagnetic relaxation. Sa Furthermore, the absence of an extension of R1 indicates that there is no increase in rotational correlation time, and Therefore, the size of the Gd: MPD-DTPA-dermatan sulfate molecular complex is relatively small ( Perhaps less than about 50,000 to 60,000 daltons). This Relatively intact (anatomically and filterably) parts of the neovascular endothelium of the tumor High tumor accessibility, even passing through in minutes, and also very fast Further evidence for surprising and unexpected benefits of high kidney clearance Is done. Both of these are observed in intact animals (see below). This result The absence of molecular aggregates detectable by laser light scattering (see above) ). The surprisingly high R1 of this new formulation is repeated multiple times and MP Gd having a D side group: DTPA (R1 = about 4 [mmol.sec]^-1New Gd in relation to : Enhancement of MPD-DTPA conjugate (and also for complete dermatan sulfate product) Seems to correlate with the observed water diffusion. Gd: Stability Kd of MPD-DTPA is 10¹⁷Exceeds .                                 Example 24   Gadolinium: MPD-DTPA: Acute mouse toxicity of ion-pair preparations of dermatan sulfate   One of the formulations of Example 22, Gd: MPD-DTPA: dermatan sulfate (60: 40% by weight) Gd: MPD-DTPA versus dermatan sulfate; 435 type dermatan sulfate, Opocrin) Gram of male Balb / C mice was tested for acute toxicity by intravenous injection into the tail vein. Tested (n = 6). Mean (Gd and chelator) when injections are given for 10-12 minutes The LD50 is 11.0 mmol / kg, with 3 mice surviving on average 9.9 mmol / kg, and Three mice died at an average of 12.2 mmol / kg. Injects more often at 2-3 minute intervals When performed rapidly, the LD50 was modestly low in dose. this Compare these results with those of Gd: DTPA (dimeglumine) (LD50 = 4.0 mmol / kg) And is advantageous.                                 Example 25     67Ga-labeled deferoxamine: dermatan sulfate; and                   111In labeled MPD-DTPA: Dermatan sulfate               Acute blood clearance of radiolabeled ion-pair preparations   Dermatan sulfate carrier has its own very fast and complete blood clearance Evaluate whether it can be given to active substances with added properties (including non-covalent chelates) To do this, the formulations of Examples 2, 5, 21, and 22 (described above) were Radioactive single proton emission (SPECT) instead of Fe (III) or Gd (III) It is modified to bind to 67Ga or 111In, which is a metal.   For experiments at 67Ga, approximately 1.55 μmol of deferoxamine (DFo) -dermatan Label sulfuric acid (77.5: 22.% wt%; DS type 435, Opocrin) with about 800 μCi of 67Ga I do. This converts 67Ga from chloride form to citrate form and has a pH of 5.5. Incubate this with DFo: dermatan sulfate for 10 minutes at room temperature at ~ 6.5 This is done by: 0.39 μmol of D in the tail vein of rats of Copenhagen strain Fo: Intravenous injection of dermatan sulfate (about 200μCi of 67Ga is chelated) A series of gamma cameras over 1 hour (and again at 24 and 48 hours) Images, and blood clearance, liver clearance and kidney clearance Heart, epigastric region, and pelvis, which are the regions of interest (ROI) Analyze the area. Average blood clearance t1 / 2 is 18 minutes, very fast 8 minutes It has a t1 / 2α component and a t1 / 2β component of 35 minutes. Liver clearance is not observed at all Not. Kidney clearance is very rapid and all recognizable clearance And lead to rapid bladder activity. Skeletal axis or region of nose, bone or bone marrow There is no significant residual activity in the region. In the control experiment, 67GaDFo Injection alone (without dermatan sulfate) is also very rapid But a significant fraction of the drug (about 30%) is very quickly (10-30 minutes) Purifies the gut and intestine, producing a high organ background in the liver and colon I will.   In another experiment (here, Copenhagen rats were implanted with AT-1 prostate implanted posteriorly In adenocarcinoma of the lung (with a diameter of 1.0-4.5 cm), the tumors are very Quickly (approximately 5 minutes) active (brighter) and tumor counts per pixel Exceeded tumor counts in blood and liver from 15 minutes after injection, Tumors are detected quickly and with high sensitivity. This means that MRI images in the same tumor model The results of the image formation are confirmed (Example 19).   MRI dose, dose escalation, and potential therapeutic dose clearance half-life In another embodiment, to assess the effect of dermatan sulfate, The amount is increased 100-fold from 1.55 μmol / kg to 155 μmol / kg (0.155 mmol / kg) The dose of the radionuclide is kept constant at 200 μCi per rat. By visual assessment , The clearance is almost identical to the drug at 100 times lower dose (described above), With only a slight (about 5 minutes) extension.   In yet another experiment, 111In was converted to the acetate form at pH 5.5-6.5 and MPD-D TPA: dermatan sulfate (60:40 wt% MPD-DTPA: dermatan sulfate, 435 type, Op ocrin) for radiolabelling. Clearance time and organ clearer The transfer pattern (kidney vs. liver) is 67GaDFo: clearer of dermatan sulfate (described above). Comparable to immunization time and tissue clearance pattern; and, when tested, Uptake is also rapid and clear.   These surprising and unexpected benefits: (a) non-covalent binding to dermatan sulfate 100 times (or better) for two different active substances that combine (by counterion binding) Or more) very rapid clearance over dose escalation; and (b) Liver and intestinal clearance is avoided in the presence of dermatan sulfate carrier Not be avoided in the absence of dermatan sulphate, Low MRI and radiation in the liver and mid-belly, which are important and difficult areas of the body It offers the major advantage of a radionuclide imaging background. Bladder catheter The pelvic area is also observed without substantial background interference . In addition, a major dose increase of at least two orders of magnitude increases blood and body clearance. Lance times only increase very slowly, meaningful therapeutic regimens Is possible. These clearance properties are consistent with the choices shown in this example and above. Selectivity and the rest of the body and all Further surprising and unexpected to enhance the difference between physical toxicity Provides even benefits.                                 Example 26     Gadolinium: N-methyl-1,3-, propanediamine-DTPA: dermatan sulfate                     (Gd: MPD-DTPA: DS) Selective contrast agent:        MRI imaging of lactating mammary adenocarcinoma in syngeneic Fisher 344 female rats   25A, 25B, 25C, 25D, 25E, and 25F. Has syngeneic breast adenocarcinoma inoculated into the liver with a trocar as in Example 18 (above) Female 344 female rats were given a dose of 0.155 mmol / kg Gd: MPD-DTPA: DS (DS = 435 Opocrin) before (pre) and after (iv) intravenous injection (i.v.). T1 weighted (T1-weighted) MRI image (TR-TE = 800/45) at 1.0 Tesla . To identify the approximate location of the tumor nodule, a T2 scout image (TR / TE = 2100/85 ) Before the T1 image contrast series (FIG. 18A). This allows two solid molds Tumor nodules (right back of liver) and one irregular tumor infiltrate (middle area of liver) And all tumor sites are subsequently confirmed by gross visual endoscopy. this These nodules cannot be identified in the T1 (800/45) pre-contrast (pre) image (Fig. 18B) After injection of Gd: MPD-DTPA: DS, all three tumor nodules are: (a) After injection It is rapidly and very strongly enhanced as early as 7 minutes (FIG. 18C); Prolonged (over 60 minutes) sharp tumor borders to the surrounding tumor-free liver (FIGS. 18C, 18D, 18E) and extended for 60 minutes ( (Persisted) (FIG. 18F) and at the tumor border 60 minutes after injection (MPI). The contrast gradient is only slightly impaired. In this animal model, Gd: MPD-DTPA: DS enhanced MRI contrast caused by DS Samin (ferrioxamine): much larger than that caused by deltaman sulfate drugs (More powerful on a dose basis); and Gd: DTPA-lysine: deltaman sulfate (implemented Prepared by Examples 3, 4, and 9; FIGS. 13A, 13B, 13C for relative intensities , FIG. 13D, Example 21, and Table 2). To moderately large (more potent on a dose basis); both of the aforementioned drugs Gd: MPD-DTA: DS (7.8 [mmol.sec]^-1Weaker metal chelate compared to R1) (Ie, having R1 of 1.6 and 4.2, respectively). The image of this embodiment is also Gd : DTPA (dimeglumine) and all reported liver-specific T1 and T2 structures Shows all of the following surprising and unexpected advantages over contrast agents: (a) The tumor itself without substantial uptake by the surrounding unrelated liver (B) enhanced tumor selectivity and sensitivity; (c) enhanced contrast Multiple sites and sites without weakening or the need for multiple contrast media injections And multiple images are acquired for improved clinical flexibility Prolonged and simultaneous immediate tumor uptake; (d) improved tumor staging ( staging) and improved at the tumor border for improved detection of small tumors Improved contrast sharpness and brightness gradient; (e) improved detection of small transitions; And (f) for enhanced prognostic and therapeutic monitoring information; Improved detection of small invasive tumor growth. Gd: MPD-DTPA: Optimal T1 image formation for DS of Surround normal liver at all indicated and suitable post-contrast times Note that there is a slight blood pool enhancement in. This is 0.15 Strongly suggests that even doses lower than 5 mmol / kg are very effective You. This compares with the Gd: MPD-DTPA chelate compared to all others described herein. Is substantially stronger [R1 = 7.8 (mmol.sec)^-1] And therefore the tumor Higher T2 per μmol of drug in tumor^*Brightness reduction and T1 brightness increase effect (For example, see Example 26 for confirmation of this effect. When).                                 Example 27     Gadolinium: N-methyl 1-1,3, propanediamine-DTPA: Dermatan sulfate     (Gd: MPD-DTPA: DS) Selective contrast agent: Syngeneic Copenhagen rat                   Imaging of prostate AT-1 adenocarcinoma in children;                       Positive correlation with special histochemical staining   As shown in FIGS.20A, 20B, 20C, 20D, and 20E, T1-enhanced imaging (T R / Te = 250/80) with Gd: MPD-DTPA: DS as prepared in Examples 21 and 22 (DS = 435 type, Opocrin) selective contrast agent at 0.155 mmol / Kg [Gd (III) dose] AT-1 prostate adenocarcinoma grown in the skin sac of syngeneic Copenhagen rats (described in Example 18). 4) before (Pre) and after (Post) injection of intravenous (I.V.) Perform at 7 Tesla. Gd: MPD-DTPA: DS was administered 7 minutes (FIG. 20B) and 20 minutes (FIG. 20B) after injection (MPI). (C) rapid, very strong T1 contrast enhancement of all tumors, as well as 40 MPI (Figure 20D) and And 60 MPI (FIG.20E), specifically the tumor margin, more particularly the basal tumor margin (where the representative Continuous strong contrast of the tumor Generate enhancements. Further experimental evaluation showed that at 40 and 60 MPI in the central tumor area The apparent moderate contrast loss of brightness that appears is due to the presence of drug in these tumor areas.Overconcentration Accurately and competes with strong T1 brightness-enhancing effects in these central tumor areas T2, which artificially reduces the brightness of the T1 contrast^*Lead the effect. This T2^*The man-made object is subjected to T / TE = 2500/250 T2 pulse sequence (T2^*Selective for effect Sensitive), and a substantially delayed post-contrast time It is detected and evaluated by observing the decrease in contrast brightness. Therefore, Gd : Very high R1 of MPD-DTPA: DS (compared to all previous drugs) with 0.155 mmol / Kg injection Very significant tumor uptake and 4.7 Tesla in combination with Gd: MPD-DT with paramagnetic response characteristics of TR / TE = 250/80 pulse sequence in the field PA: As DS accumulates over time, excessively in tumors, especially in the central area of the tumor Provides high local paramagnetic activity. The margin, especially the basal margin, is where the drug This T2 is used for more rapid back diffusion into plasma^*Relatively protected from brightness reduction artifacts Is done. The previous results and discussion indicate that doses lower than 0.155 mmol / Kg may result in Gd: MPD-DTPA: It leads to the conclusion that it is shown for optimal T1 imaging using DS. Because Gd : MPD-DTPA chelate is substantially stronger than all others described herein It is a powerful T1 paramagnetic active substance. Unrelated liver around three liver tumor nodules In the blood pool background, which is very little increased in Note Example 25, which appears to be a slight overdose, as evidenced. And also Regardless, these measures still retain all post-contrast time (7-60 MPI ) Are exceptionally well visualized.   Microwave-enhanced pull of these MRI images and Gd (III) metal ions Correlation with cyan blue staining was excised at 60 PMI (and new for sectioning and staining). Gd: MPD-DTPA: DS Gd (III) that becomes localized on the outer 2/3 of the tumor mass (freshly frozen) (As described in Example 18). (See FIG. 20). This is almost Showing strongly positive histochemical staining of all tumor cells and a significant number of tumor cells The cells have a positive nuclear staining (ie, nuclear localization of the metal ion marker). Yo Less number of (breast) tumor cells stained lighter in 7 minutes (Example 18) Almost all tumor cells stained very strongly in 60 minutes, and in this example At 60 minutes but not in (breast) tumors at 7 minutes (Example 18) Further nuclear localization The internalization is that the internalization of the tumor cells is at one hour intervals, perhaps at full intervals of time (during which time, Dermatan sulfate-bound metal chelate maintained at significant concentrations in extracellular matrix Extremely rapid and selective extravasation across tumor-induced angiogenic MRI endothelium (Fills first and quickly through local microvessels) Strongly suggests-including tumor selective induction, and VEGF / VPF plus others See text above regarding endothelial localization of GAG-coupled receptors. Pre-malignant Surprising and unexpected benefits of endothelial localization observed here for gonad tumors Was also observed in Example 18 for malignant breast tumors. This is the The surprising and unexpected findings in Example 18 were confirmed, where within tumor-induced angiogenesis The skin and the appropriate tumor cells are conjugated to the MRI contrast agent dermatan sulfate Class of binding, pumping, extravasation and tumor cell internalization Target and, in fact, dermatan sulfate and other activities that bound to GAG as well It is a drug target. Localization of tumor endothelium, tumor matrix, tumor cells and nucleus These findings of accumulation and furthermore, metal chelates or other types of active substances Provide the basis for selective localization of the therapeutic agent, regardless of whether                                 Example 28        As an ion-pair complex with essentially purified dermatan sulfate                         Preparation of doxorubicin preparation   Intrinsically purified dermatan sulfate (435 type, Opocrin, formal MW = 18,00 (0 Daltons) in water at 10 mg / ml and high purity of 4 mg / ml with vigorous stirring Solution of doxorubicin (Meiji Seika Kaisha, Ltd., Japan) was added dropwise at a ratio of 60: 4. 0 (w / w) of doxorubicin versus dermatan sulfate is obtained. (Between 10:90 and 90:10 (w / w) Other ratios of doxorubicin to dermatan sulfate are also tested). The mixture 8 minutes by ultrasonic wave at 4 ° C. using a macro probe sonicator (Heat Systems) Homogenize for a while. This is a laser light scattering doxorubicin: dermatan sulfate complex Its limit of 11 nanometers (small as assessed by the Nicomp system) ) Effectively reduce to size. The obtained liquid is filtered with a 0.22 μm low binding filter ( Filtration through a Millipore, Millex GV), 500 mg / ml sucrose solution (Boehringer Man nheim) was added, stirred and then 10 mg / mL polyethylene glycol (Hoechs t, average MW = 3,350 daltons) and add the resulting solution to ultrasound again. Run (as above) and filter again with a 0.22um GV filter for sterility And fill vials and store as liquid or frozen, and place on suitable shelves And Lyophilize for 17 hours primary drying cycle at room temperature and conditions. To obtain a well-formed red brick-colored cake with a residual moisture of about 2.0% (Karl F isher method). Plug and seal the vial. For use, cake is sterile water at 2mg / ml (Shake by hand for 15 seconds). The resulting liquid is completely translucent, red-red Yellow, pH 6.8-7.1, osmolality of about 210 mOsm / Kg, and -38 It has a ζ potential of −−40 mV, indicating that dermatan strongly bound in a slight molar excess The presence of sulfuric acid (doxorubicin itself has its sugar amine group (this is a strong ion-pair Is effective in binding sulphate of dermatan sulphate). ζ potential). The freeze-dried cake is prepared with (Doxorubicin and Derma (Doxorubicin) are stable at room temperature and at 4 ° C for long periods of time. HPLC analysis: C-18 Lichrosphere; mobile phase: acetonitrile: water (50:50) ), 20 mM phosphoric acid + 5 mM sodium dodecyl sulfate, pH 2.3; HPLC of dermatan sulfate Analysis by: TSK molecular sieve; mobile phase: 0.2 M sodium sulfate for charge suppression, 1 , 800 to 17,250 daltons with Opocrin dermatan molecular weight standards). Reconstructed Meet the USP specifications for oversized particles of 10um and 25um and larger (Hyac-Royco By laser analysis).   Adriamycin (a source of Adria Laboratories doxorubicin) is also available from Dell. Prepared similarly by ion-pairing with matane sulfate (Opocrin, supra).                                 Example 29     Preparation of doxorubicin formulation as ion-pair complex with bovine lung heparin   High-purity doxorubicin solubilized bovine lung heparin (Hepar-Kabi-Pharmacia) And a ratio ranging from 90:10 to 10:90 (w / w, doxorubicin: heparin) and one Mix at the optimal ratio, i.e. 60:40 (w / w), then mix as described in Example 28. And subject to further steps. Obtains a visually clear, 0.22um filterable liquid You. However, this liquid has a larger limit of 25 nanometers due to laser light scattering. Degree size; and the lyophilized cake is for rapid homogenous reconstitution And is considered more resistant (requires about 1-2 hours).                                 Example 30         With essentially purified dermatan sulfate and bovine lung heparin                 Preparation of coated taxol nanoparticle formulation   These formulations are treated in two steps, first with taxol being lecithin or lecithin- By solubilizing and preparing in a sterol nanodispersion, and Dermatan sulfate (435 type, Opocrin) with nanodispersion coated with lecithin on eyes Or prepared by interacting with bovine lung heparin (Hepar-Kaki-Pharmacia) And the binding of glycosaminoglycan sulfate to the highly basic nitrogen group of lecithin In both cases, the final nanodispersion is formed and formed by ion-pair interaction at the nanoparticle surface And stabilize. Taxol (Sigma Chemical Co., St. Louis) was converted to methylene chloride And yolk lecithin into chloroform or soy lecithin + cholesterol Dissolve in methylene chloride. Place the solubilized components in a round bottom flask and add solvent Evaporate under vacuum for 30 minutes to remove taxol lecithin (in some cases, cholesterol). Is also formed). After drying thoroughly, water (under nitrogen) is added and Hydrate the probe for 2 minutes at 4 ° C. (Heat Syst. em). Dermatan sulfate, or alternatively bovine lung heparin, at 2-6% (w / w, And the mixture probe is sonicated for 1 minute at 4 ° C. Obtained Observed nanoparticles with an optical microscope to detect surface glycosaminoglycan (GAG) Is confirmed by the addition of the soluble dye Azure A, which rapidly binds to the surface GAG Turns purple and produces particle aggregation (lecithin only, uncoated Preparation = negative). Coated with dermatan sulfate coated nanoparticles and heparin Microfiltration and solubilization of the size and amount of drug in the Taxol was evaluated by UV absorption analysis (230 nm) and sized. Furthermore, it is confirmed by a laser light scattering method (Nicomp). Contains 4% glycosaminoglycan The formulation with is optimal. Present before filtration ("none"), and various pore sizes (5.0, 0.45 um and 0.22 um) for the amount of taxol remaining after filtration. Tabular results are shown in Table 3 for the dermatan sulfate formulation.   The results of these analyzes indicate that both lecithins were dermatan sulfate and bovine lung hepain. Glycos that effectively interact with phosphorus and stabilize taxol nanoparticle dispersions Figure 4 shows forming a Saminoglycan surface coating. However, the best of lecithin For the appropriate type (yolk), the resulting nanoparticle size (laser light scattering (Table 4 ), Heparin (column) for dermatan sulfate (column a). c) and only dermatan sulfate converts taxol to sterile Formulate with acceptable recovery rate through a cut-off filter (0.45 um), filter, and To obtain a standard taxol solubilizer, Cremophor without the need for (cremofor), and its inherent toxicity and acute allergic side effects Provides a sterile, intravenously acceptable taxol nanodispersion without having .   This smaller nanoparticle size for dermatan sulfate versus bovine lung heparin is Similar to the size observed for doxorubicin (Example 28). this is, Further surprising and unexpected formulations of dermatan sulfate against heparin Check the benefits (above).                                 Example 31           Has essentially purified dermatan sulfate (435 type)                     Preparation of vincristine ion pair preparation   Vincristine (Sigma Chemical Co., St. Louis) is dissolved in water and 90:10 And dermatan sulfate at a ratio of drug to dermatan sulfate between 30:70 (w / w) You. The result is a clear solution, with the optimal ratio present at 30-40% (w / w) of the drug. No particles are detected (by laser light scattering). The result is a 500 MW cut-off. Combined with the retention of the ion pair form (rather than the drug alone) inside the dialysis bag A strong ion pair is formed between the amine group of lincristine and the sulfate group of dermatan sulfate. It shows that                                 Example 32 With essentially purified dermatan sulfate (435 type) and bovine lung heparin Amine-containing antibiotic anti-infectives, amikacin, gentamicin,       Of Tobramycin and Tobramycin as Ion Pair Preparations   Amikacin, gentamicin, and tobramycin (all Sigma Chemical C o., St. (Obtained from Louis) in water, and the drug versus glycosaminoglycan Dermatan sulfate (435 type) or in a ratio between 90:10 and 30:70 (w / w) or Mix with bovine lung heparin (Hepar-Kabi-Pharmacia). 30-50% of the optimal range (w / w), a strong ion-pair complex is formed between the drug and glycosaminoglycan I do. This is due to laser light scattering (Nicomp) (100-2 for bovine lung heparin). 00 nanometer range) or smaller dermatan sulfate formulation (laser light scattering 500 MW cut-off dialysis retention) Will be revealed.                                 Example 33             Basic peptide and essentially purified dermatan sulfate                       (435 type) and ion pair preparation   The following basic leukocyte chemoattractants and inflammatory peptides: (a) N-formyl-m et-leu-phe-lys (acetate), (b) arginine bradykinin, (arg-pro-pr o-gly-phe-ser-pro-phe-arg) and (c) poly-L-lysine (all three are Sigma)  Chemical Co., St. Dissolve from water) and 90:10 to 10:90 (active Essentially purified dermatan sulfate (435% w / w of dermatan sulfate). Mix with type, Opcrin). Strong ion-pair binding is optimized for the optimal ratio of about 60:40 (active Dermatan sulfate). This is due to laser light scattering, 500 MW Retention in toof dialysis bags, and (a) in the case of N-formyl-met-leu-phe-lys , Revealed by increased fluorescence at an emission wavelength of 518 nm (excitation wavelength = 257 nm) .   These basic peptides in a formulation with essentially purified dermatan sulfate , For the purpose of local treatment (due to significant systemic inflammatory side effects, free To recover endogenous or infused leukocytes (under conditions where the drug cannot be tolerated) Biomodulating peptides at the site of tumor and / or infection to activate Provides a novel means for site-selective localization, accumulation, retention, and action of tides You. Therefore, these new dermatan sulfate formulations are useful in tumor therapy and in vivo. It offers surprising and unexpected benefits to anti-infective and anti-infective therapies.                                 Example 34           Against wild-type and doxorubicin-resistant human breast cancer cells       Ion-pair doxorubicin: dermatan sulfate formulation for comparative activity                 In vitro testing of standard and standard doxorubicin   Tumor cell killing ability in clonogenic assay The ion pair doxorubicin: dermatan sulfate of Example 28 (essentially purified Dermatan sulfate, 435 type, Opocrin) (= doxorubicin: DS) as standard doxo Compare with Rubicin solution (Adria Laboratories). This comparison includes the following human milk Cell line: MCF-7 cell line parent and adriamycin / doxorubicin Resistant MCF-7 cell line. For each group, make 5 serial dilutions of each formulation on day 0 The appropriate number of cells, growth medium, and¹⁴Mix with C-labeled glucose and then Premix 5% CO2_TwoInject into a gas-filled serum vial and place the vial in 3 Incubated at 7 ° C. (cells were continuously exposed to drug). Bactec machine (Bactec Corp.) on days 6, 9, and 12 of the incubation. Produced in control and drug treated vials¹⁴C glucose CO_TwoAmount of And record the data as% survival, IC50 and IC90 values, and When analyzed, the% survival values were determined for control cells (incubated without drug). ) Is 100% larger than the standardized value of 100. Day of peak count (9 The results for (day) are shown in Table 5.   MCF-7 parent strains have similarly low IC50 and IC90 concentrations, but doxorubicin-resistant M In the CF-7 strain, the IC50 and IC90 concentrations are extremely different, and doxorubicin-del Matan sulfate can overcome (or circumvent) resistance, whereas standard doxorubicin does The fact that dermatan sulfate cannot be used (or avoided) indicates that Overcoming the phenotype, which is to avoid the Pgp (P-glycoprotein) pump It is strongly suggested that this can be done. This is due to the effective negative charge of dermatan sulfate. Can be predicted from the load. Despite the specific mechanism, this result: Dermatan sulfate antitumor preparations in the treatment of tumors and neoplastic diseases, in particular Sorubicin dermatan doxorubicin sulfate, most specifically, essentially purified Further important and surprising of doxorubicin dermatan sulfate formulation (Example 28) And provide unexpected benefits. These test assays are based on Cancer Therapy a nd Research Center, Institute for Drug Development, San Antonio, Texas Donna Degen, M.S. and Daniel D. To what Von Hoff, M.D. Please note.                                 Example 35      Ion-pair doxorubicin: Acute in vivo toxicity test of dermatan sulfate   Male Balb / c mice (n = 4 / group) were treated with the ion pair doxorubici of Example 28. Dermatan sulfate (essentially purified dermatan sulfate, type 435, Opocrin ) (= Doxorubicin: DS) or standard doxorubicin solution (Adria Laboratories) ) Is injected intravenously at 22.5 mg / Kg, and then housed in a cage with a filter stopper. And observe until the day of death. 22.5 mg / Kg is the standard for doxorubicin Is the LD90 reported in the mouse and is necessary to observe the test endpoint. Selected to minimize time; at lower doses, current protocol Note that any differences that can be observed in are expected to be large. Day of death : Doxorubicin: DS: Model date = 6, average 5.5 ± 0.9 SE; Standard doxorubicin : Model date = 5, average 4.8 ± 0.5 SE. Therefore, doxorubicin is Tend to have acute toxicity of at least comparable to and beyond standard doxorubicin , But the differences in this test are not statistically significant. As a result Has the potential to overcome adriamycin / doxorubicin resistance in human tumor cells Combining the advantages of point (Example 34) and more effective localization in vivo , At fixed doses, in animal tumors and in tumor cell sites (see Example 36 below). Dermatan sulfate antitumor in the treatment of tumors and neoplastic diseases Drugs, specifically doxorubicin dermatan doxorubicin sulfate, and essentially purified Further surprising of the prepared doxorubicin dermatan sulfate formulation (Example 28) Provides more unexpected benefits.                                 Example 36    Ion-pair doxorubicin compared to standard doxorubicin: dermatan sulfate       In vivo tumor localization of tumors and tumor cell internalization   Example 28 Ion pair doxorubicin: dermatan sulfate (essentially purified Rumatan sulfate, 435 type, Opocrin) (= doxorubicin: DS) and standard doki Sorubicin solution (Adria Laboratories) with 5 mg / Kg of doxorubicin, subcutaneous capsule (s Copenhagen rats with AT-1 prostate cancer growing in kin pouch (Similar to growth) was injected i.v. Rats were sacrificed 3 hours after injection and tumor and Removing the major organ, placing the cut tumor and organ pieces in the OCT polymer, and Freeze at 4 ° C. and cut cryostat sections at 8 μm and cover Place Rhodamine type bandpass filter (select doxorubicin (At about 485 nm to excite) and directly the fluorescence of doxorubicin (530 Fluorescence microscopy was performed by exciting the sections to be evaluated (at emission wavelengths greater than And determine the following:   ◆ Relative tumor drug levels;   ◆ at the proximal site of tumor microvessels and at the site more distal from tumor microvessels, Depth and homogeneity of drug penetration into the tumor mass;   ◆ tumor targets, ie, endothelial cells and tumor cells in a strict sense;   ◆ Normal organ fluorescence (as a predictor of clearance and potential toxicity) (Table 6).   Doxorubicin: See Figure 21A for DS-Confluent sheet of tumor cells But almost all tumor cells (= tumor cell internalization) and neogenesis It has very bright fluorescence in the vascular endothelium. Doxorubicin: DS Figure 21 See B-More flexible clusters of tumor cells on endothelial stalk . The more flexible cluster of tumor cells (Fig. It appears to be growing or dividing (compared to the sheet). Almost all cells Is extremely brightly stained, plus the current location in this site and in the cytoplasm. Note the significantly brighter nuclear fluorescence of doxorubicin. In addition, intradermal cells and Also note the strong fluorescence of the nuclei of the intradermal cells. Standard doxorubicin (fine Vesicle (See Figure 21C for a denser sheet and a more flexible cluster in the lower left) Thing-all have significantly lower fluorescence, and within and around tumor microvessels (Center of image) as well as generally lack fluorescence in the nuclei of tumor cells.   The fluorescence intensity and pattern in other major organs was determined by the doxorubicin: DS Alance shifts away from the kidney (compared to standard doxorubicin), and Access to bile via the liver in an accelerated manner (compared to standard doxorubicin) Indicates that it causes purification. In the cardiac pulp or red spleen No increase was observed (such an increase could be expected toxicity at these sites, and These sites are the major sites of toxicity for standard doxorubicin), and The fluorescence levels at these two sites are rather comparable to standard doxorubicin. Doxorubicin: slightly lower for DS than for doxorubicin.   These results indicate that the drug (not just doxorubicin) : Significantly higher tumor localization, extracellular matrix when formulated as DS Permeation depth and width, internalization of tumor cells Doxorubicin nucleus Internal migration (the major intracellular site of action of doxorubicin) and these results indicate In addition, the induced tumor endothelium and endothelial nucleus (for doxorubicin: DS) A surprising and unexpected uptake induced by These amazing and An unexpected advantage is the advantage associated with the essentially purified dermatan sulfate of the present invention. If clearly, completely and essentially purified (along with the advantages of the previous examples (29-35)) Formulations of the invention of doxorubicin with dermatan sulfate and other cancers Distinguish from cancer therapeutic (and diagnostic) actives.

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Claims (1)

Claims: A drug carrier comprising a drug in combination with essentially purified dermatan sulfate having a sulfur content of up to 1.9% (w / w) and selective oligosaccharide oversulfation. A composition, wherein the composition has a non-occlusive size of less than 500 nm. 2. The drug carrier composition according to claim 1, wherein the carrier has a size of less than 250nm, preferably less than 25nm. 3. 2. The drug carrier composition of claim 1, wherein binding to the disease-inducing endothelium causes the endothelium to completely or partially cover the bound drug carrier composition in less than 10-15 minutes. . Said essentially purified dermatan sulfate having sulfur content and selective oligosaccharides of over sulfated up 4.9% (w / w), contains Ido-GalNAc4SO _3, and IdoA2SO ₃ -GalNAc4SO further containing ₃ and IdoAGalNAc4,6SO _3, drug carrier composition of claim 1. 5. 2. The drug carrier composition of claim 1, further defined by being a nanoparticle. 6. 2. The drug carrier composition according to claim 1, wherein the drug is a tumor therapeutic. 7. The drug carrier composition according to claim 6, wherein the tumor therapeutic agent is selected from the group consisting of adriamycin, doxorubicin, epirubicin, daunorubicin, and idarubicin, or salts thereof. 8. The tumor therapeutic agent is bleomycin, taxol, taxotere, vinblastine and vincristine, amsacrine, azacitidine, dideoxyinosine, dihydro-5-azacytidine, ethanidazole, etiophos, methotrexate, misonizadol, porphyromycin, pyrazolo acridinez, terephthalamide 7. The drug carrier composition according to claim 6, wherein the drug carrier composition is selected from the group consisting of taxotere, topotecan, trimetrexate, and carboplatin, or salts thereof. 9. 2. The drug carrier composition according to claim 1, wherein the drug is a chelator. 10. 2. The drug carrier composition according to claim 1, wherein the drug is an anti-infective. 11. The drug carrier composition according to claim 10, wherein the drug is gentamicin, tobramycin, or amikacin. 12. 2. The drug carrier composition according to claim 1, wherein the drug is a biological response modifier. 13. The drug carrier composition according to claim 1, wherein the drug is a biologically active peptide or polypeptide. 14． 14. The drug carrier composition according to claim 13, wherein said biologically active peptide or polypeptide is selected from the group consisting of leukocyte chemoattractant, bradykinin, and poly-L-lysine. 15. 15. The drug carrier composition according to claim 14, wherein the leukocyte chemoattractant is N-formyl-met-leu-phe-lys (SEQ ID NO: 1). 16. 2. The drug carrier composition of claim 1, further defined by being in a pharmaceutically acceptable solution suitable for intravascular or other parenteral injection. 17． 2. The drug carrier composition of claim 1, wherein the combination is formed by an ion-paired, amphoteric, or hydrophobic interaction between the carrier and a drug. Use of a drug carrier composition containing a tumor therapeutic in combination with essentially purified dermatan sulfate with a sulfur content of up to 18.9% (w / w) and selective oligosaccharide oversulfation Wherein the carrier has a non-occlusive size of 500 nm or less and the drug carrier composition is pharmaceutically acceptable for the manufacture of a medicament for the treatment of a tumor responsive to a tumor therapeutic. Use contained in a carrier. Comprises 19.9% (w / w) the essentially purified dermatan sulfate having sulfur content and selective oligosaccharides of over sulfated until the Ido-GalNAc4SO _3, and IdoA2SO ₃ -GalNAc4SO ₃ and further comprising IdoAGalNAc4,6SO _3, use according to claim 18. 20. 19. The use according to claim 18, wherein the drug carrier composition is further defined by being nanoparticles. 21. 19. The use according to claim 18, wherein the tumor therapeutic is selected from the group consisting of adriamycin, doxorubicin, epirubicin, daunorubicin, idarubicin, bleomycin, taxol, taxotere, vinblastine, and vincristine, or salts thereof. 22. The tumor therapeutic agent is amsacrine, azacytidine, dideoxyinosine, dihydro-5-azacytidine, ethanidazole, etiophos, methotrexate, misonizador, porphyromycin, pyrazolo acrizinex, terephthalamidine, topotecan, trimetrexate, and carboplatin, or 19. The use according to claim 18, wherein the use is selected from the group consisting of salts thereof. 23. 19. Use according to claim 18, wherein the tumor therapeutic is in a controlled release form. 24. 19. The method of claim 18, wherein binding of the sample of the drug carrier composition to disease-induced endothelium results in endothelial induction to completely or partially cover the bound sample within less than 10-15 minutes. use. 25. 19. The method of claim 18, wherein the drug carrier composition is for the manufacture of a medicament to be administered by selected arterial perfusion to obtain high efficiency uptake in a proximal target organ, tissue, or tissue lesion. use. 26. 19. The use according to claim 18, wherein the drug carrier composition is for the manufacture of a medicament to be administered intravenously to obtain semiselective uptake in tissue lesions located at widely distributed systemic sites. .