# US 5,707,604 A · Vivo agents comprising metal-ion chelates with acidic saccharides and glycosaminoglycans, giving improved site-selective localization, uptake mechanism, sensitivity and kinetic-spatial profiles

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 5,707,604 A
- Document ID: US5707604A
- Status: Granted
- Kind: A
- Application number: 08/472,634
- Filed: 1995-06-07
- Granted: 1998-01-13
- Earliest claimed: 1986-11-18
- Assignees: Access Pharmaceuticals, Inc.
- Inventors: David F. Ranney
- Web page: https://patentgrep.com/patent/US5707604A

## CPC

- C08B 37/0063: {Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan}
- B82Y 5/00: Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
- Y10T 436/24: Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry
- C08B: POLYSACCHARIDES; DERIVATIVES THEREOF (polysaccharides containing less than six saccharide radicals attached to each other by glycosidic linkages C07H; fermentation or enzyme-using processes C12P19/00; sugar industry C13; production of cellulose D21)
- B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

This application concerns novel agents comprising cationic or chemically basic metal chelators in association with hydrophilic carriers of anionic or chemically acidic saccharides, sulfatoids and glycosaminoglycans. In certain embodiments, the agents comprise metals and metal ions. Covalent and non-covalent chemical and physical means are described for stabilizing the binding of the metal chelators to the carriers. Novel non-covalently bound compositions are described which give uniquely high payloads and ratio of metal chelator to carrier, ranging from a low of about 15% metal chelator by weight, to a characteristic range of 70% to 90% metal chelator by weight. Specific embodiments are described comprising deferoxamine, ferrioxamine, iron-basic porphine, iron-triethylenetetraamine, gadolinium DTPA-lysine, gadolinium DOTA-lysine and gadolinium with basic derivatives of porphyrins, porphines, expanded porphyrins, Texaphyrins and sapphyrins as the basic or cationic metal chelators, which are in turn, bound to acidic or anionic carriers, including one or more of acidic or anionic saccharides, and including sulfated sucrose, pentosan polysulfate, dermatan sulfate, essentially purified dermatan sulfate with a sulfur content of up to 9% and with selective oligosaccharide oversulfation, chondroitin sulfate, oversulfated chondroitin sulfate, heparan sulfate, beef heparin, porcine heparin, non-anticoagulant heparins, and other native and modified acidic saccharides and glycosaminoglycans.

## Claims

### Claim 1

An agent comprising a metal-ion chelate, said chelate having at least one excess basic or cationic group not involved in metal-ion chelation and being bound to an anionic, hydrophilic, water-soluble glycosaminoglycan carrier with selective saccharide oversulfation and a sulfur content between about 4% (w/w) and about 9% (w/w).

### Claim 2

An agent comprising a metal-ion chelate, said chelate having at least one excess basic or cationic group not involved in metal-ion chelation being noncovalently bound to an anionic, hydrophilic, water-soluble glycosaminoglycan carrier with selective saccharide oversulfation and a sulfur content between about 4% (w/w) and about 9% (w/w) by paired ion electrostatic binding.

### Claim 3

An agent comprising a metal-ion chelate, said chelate having at least one excess basic or cationic group not involved in metal-ion chelation and being covalently bound to an anionic, hydrophilic, water-soluble glycosaminoglycan carrier with selective saccharide oversulfation and a sulfur content between about 4% (w/w) and about 9% (w/w).

### Claim 4

An agent comprising a metal-ion chelate, said chelate having at least one excess basic or cationic group not involved in metal-ion chelation and being bound to essentially purified dermatan sulfate carrier with a sulfur content between about 4% (w/w) and about 9% (w/w) and with selective oligosaccharide oversulfation.

### Claim 5 (depends on 1, 2, 3, 4)

The image-enhancing agent or spectral-enhancing agent of claim 1, 2, 3 or 4 further comprising ferrioxamine covalently conjugated to the glycosaminoglycan carrier by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, or carbonyldiimidazole.

### Claim 6

A spectral-enhancing agent to enhance images arising from induced magnetic resonance signals, the agent comprising Gd(III)diethylenetriaminepentaacetate covalently conjugated to heparin or essentially purified dermatan sulfate with a sulfur content of from about 4% to about 9% (w/w) and with selective oligosaccharide oversulfation.

### Claim 7 (depends on 1, 2, 3, 4)

The agent for enhancing body imaging of claim 1, 2, 3 or 4, where the chelate is diethylenetriaminepentaacetate-lysine, and the agent comprises chelated Gd(III).

### Claim 8 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4 where the chelate further comprises chelated Gd(III) and the chelate is 1,4,7,10-tetraazacyclododecane-N,N',N:,N'"-tetraacetate-lysine (DOTA-lysine).

### Claim 9

An agent comprising ferrioxamine bound by non-covalent electrostatic binding to essentially purified dermatan sulfate with a sulfur content between about 4% (w/w and about 9% (w/w) and with selective oligosaccharide oversulfation.

### Claim 10

An agent comprising N-methyl-1,3-propanediamine-DTPA bound to essentially purified dermatan sulfate with a sulfur content between about 4% (w/w) and about 9% (w/w) and with selective oligosaccharide oversulfation.

### Claim 11

An agent comprising gadolinium (III):N-methyl-1,3-propanediamine-DTPA bound to essentially purified dermatan sulfate with a sulfur content between about 4% (w/w) and about 9% (w/w) and with selective oligosaccharide oversulfation.

### Claim 12 (depends on 4)

The agent of claim 4, wherein said essentially purified dermatan sulfate has at least about 220 U/mg heparin cofactor II activity.

### Claim 13 (depends on 4)

The agent of claim 4, wherein said essentially purified dermatan sulfate contains Ido-GalNAc4SO.sub.3 and further comprises IdoA2SO.sub.3 GalNAc4SO.sub.3 and IdoAGalNAc4,6SO.sub.3.

### Claim 14 (depends on 1, 2, 3, 4, 12, 13)

The agent of claim 1, 2, 3, 4, 12 or 13 wherein the metal-ion chelate is a paramagnetic metal-ion chelate.

### Claim 15 (depends on 1, 2, 3, 4, 12, 13)

The agent of claim 1, 2, 3, 4, 12 or 13 further defined as being at least about 15 weight percent metal-ion chelate.

### Claim 16 (depends on 1, 2, 3, 4, 12, 13)

The agent of claim 1, 2, 3, 4, 12 or 13, further defined as comprising a metal ion selected from the group consisting of iron, manganese, chromium, copper, nickel, gadolinium, erbium, europium, dysprosium and holmium.

### Claim 17 (depends on 1, 2)

The agent of claim 1, 2, or 3, wherein the glycosaminoglycan is selected from the group consisting of heparan chondroitin sulfate, glycine-conjugated heparin, heparan sulfate, and dermatan sulfate.

### Claim 18 (depends on 1, 2, 3)

The agent of claim 1, 2 or 3 wherein the glycosaminoglycan is chondroitin sulfate.

### Claim 19 (depends on 4, 11)

The agent of claim 4 or 11, wherein said essentially purified dermatan sulfate is further defined as having a molecular weight of from about 8,000 daltons to about 45,000 daltons.

### Claim 20 (depends on 4, 11)

The agent of claim 4 or 11, wherein said essentially purified dermatan sulfate is further defined as having a molecular weight of from about 10,000 daltons to about 23,000 daltons.

### Claim 21 (depends on 4, 11)

The agent of claim 4 or 11, wherein said essentially purified dermatan sulfate is further defined as having a molecular weight of from about 13,000 daltons to about 19,000 daltons.

### Claim 22 (depends on 4, 11)

The agent of claim 4 or 11, wherein said dermatan sulfate is further defined as having greater than 220 U/mg heparin cofactor II activity.

### Claim 23 (depends on 4, 11)

The agent of claim 4 or 11, wherein said dermatan sulfate is further defined as having a SO.sub.3 /COO.sup.- ratio of between 0.7:1 and 1.8:1.

### Claim 24 (depends on 4, 11)

The agent of claim 4 or 11, wherein said dermatan sulfate is further defined as having a SO.sub.3 /COO.sup.- ratio of between 0.9:1 and 1.5:1.

### Claim 25 (depends on 1, 2, 3)

The agent of claim 1, 2, 3, or 4, wherein said metal-ion chelate has a formation constant for paramagnetic metal ions of at least about 10.sup.14.

### Claim 26 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4, wherein said metal-ion chelate is an iron chelate.

### Claim 27 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4, wherein the chelator of said metal-ion chelate is a hydroxamate.

### Claim 28 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4, wherein said metal-ion chelate is ferrichrome, ferrioxamine, enterobactin, ferrimycobactin or ferrichrysin.

### Claim 29 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4, wherein said metal-ion chelate is ferrioxamine.

### Claim 30 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4 wherein said metal-ion chelate is gadolinium (III):N-methyl-1,3-propanediamine-DTPA.

### Claim 31 (depends on 1, 2)

The agent of claim 1, 2, or 3, wherein said metal-ion chelate is ferrioxamine and said glycosaminoglycan carrier is heparin or a heparin fragment.

### Claim 32 (depends on 1, 2)

The agent of claim 1, 2, or 3, wherein said metal-ion chelate is ferrioxamine and said the glycosaminoglycan carrier is essentially purified dermatan sulfate.

### Claim 33 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4, wherein said metal-ion chelate is gadolinium (III):N-methyl-1,3-propanediamine-DTPA.

### Claim 34 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4, wherein said metal-ion chelate is further defined as comprising a porphine, porphyrin, sapphyrin or texaphyrin.

### Claim 35 (depends on 34)

The agent of claim 34, further defined as comprising an iron ion or a gadolinium ion.

### Claim 36 (depends on 1, 2)

The agent of claim 1, 2, or 3, wherein the chelator of said metal-ion chelate is 5, 10, 15, 20-tetrakis(1-methyl-4-pyridyl)-21H,23H-porphine, and the agent further comprises a chelated iron ion.

### Claim 37 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4, wherein said metal-ion chelate is further defined as comprising a polyaminocarboxylate or macrocyclic compound.

### Claim 38 (depends on 37)

The agent of claim 37, wherein said polyaminocarboxylate is a basic or amine derivative of diethylenetriaminetetraacetate.

### Claim 39 (depends on 37)

The agent of claim 37 wherein said macrocyclic compound is a basic or amine derivative of 1,4,7,10-tetraazacyclododecane-N,N',N",N'"-tetraacetate (DOTA).

### Claim 40 (depends on 37)

The agent of claim 37 wherein said polyaminocarboxylate is N-methyl-1,3-propanediamine-DTPA.

### Claim 41 (depends on 1, 2, 3, 4)

The agent of claim 1, 2, 3 or 4, wherein said carrier is defined further as selectively binding endothelial determinants selectively induced at disease sites.

### Claim 42 (depends on 10)

The agent of claim 10, further comprising a gadolinium (III) ion.

### Claim 43 (depends on 1, 2, 3, 4)

The agent of claims 1, 2, 3 or 4, wherein the chelator of said metal-ion chelate comprises a diamine chemical side group or a diamine chemical side group derivative.

### Claim 44 (depends on 1, 2, 3, 4, 9, 10, 11)

The agent of claim 1, 2, 3, 4, 9, 10 or 11 defined further as being in a combination with at least one of a buffer or salt, to produce an osmotic strength suitable for parenteral administration, and as being an aqueous solution or a lyophilized or dry preparation suitable for aqueous reconstitution having the desired osmotic strength, and wherein said agent is aseptic or sterile.

### Claim 45 (depends on 44)

A method of enhancing magnetic resonance images or spectra in vertebrate animals comprising administering to said animal an effective amount of the agent of claim 44.

### Claim 46 (depends on 44)

A method of enhancing in vivo images arising from induced magnetic resonance signals, comprising the steps of:
administering to a subject an effective amount of the agent of claim 44;
exposing the subject to a magnetic field and radiofrequency pulse; and
acquiring an induced magnetic resonance signal to obtain a contrast effect.

### Claim 47 (depends on 11)

A method of preparing the agent of claim 11 comprising the steps of:
forming a solution of essentially purified dermatan sulfate with a sulfur content between about 4% (w/w) and about 9% (w/w) and with selective oligosaccharide oversulfation at a concentration of about 400 mg/ml; and
mixing with said solution gadolinium (III):N-methyl-1,3-propanediamine-DTPA.

### Claim 48 (depends on 47)

The method of claim 47, further comprising the step of filtering the agent through a 0.22 micrometer sterile filter to render the filtrate aseptic.