Lipid Construct for Delivery of Insulin to a Mammal

Abstract

The instant invention is drawn to a hepatocyte targeted composition comprising insulin associated with a lipid construct comprising an amphipathic lipid and an extended amphipathic lipid that targets the construct to a receptor displayed by an hepatocyte. The composition can comprise a mixture of free insulin and insulin associated with the complex. The composition can be modified to protect insulin and the complex from degradation. The invention also includes methods for the manufacture of the composition and loading insulin into the composition and recycling various components of the composition. Methods of treating individuals inflicted with diabetes.

1 . A lipid construct comprising an amphipathic lipid and an extended amphipathic lipid, wherein the extended amphipathic lipid comprises proximal, medial and distal moieties, wherein the proximal moiety connects the extended amphipathic lipid to the construct, the distal moiety targets the construct to a receptor displayed by a hepatocyte, and the medial moiety connects the proximal and distal moieties. 
     
     
         2 . The lipid construct of  claim 1 , further comprising at least one insulin. 
     
     
         3 . The lipid construct of  claim 2 , wherein the at least one insulin is selected from the group consisting of insulin lispro, insulin aspart, regular insulin, insulin glargine, insulin zinc, human insulin zinc extended, isophane insulin, human buffered regular insulin, insulin glulisine, recombinant human regular insulin, recombinant human insulin isophane, premixed combinations of any of the aforementioned insulins, a derivative thereof, and a combination of any of the aforementioned insulins. 
     
     
         4 . The lipid construct of  claim 2 , further comprising an insoluble form of at least one insulin associated with the lipid construct. 
     
     
         5 . The lipid construct of  claim 1 , wherein the amphipathic lipid comprises at least one lipids selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine, cholesterol, dicetyl phosphate, 1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycero)], 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl), derivatives thereof, and mixtures of any of the foregoing compounds. 
     
     
         6 . The lipid construct of  claim 1 , wherein the proximal moiety of the extended amphipathic lipid comprises at least one, but not more than two, long acyl hydrocarbon chains bound to a backbone, wherein each hydrocarbon chain is independently selected from the group consisting of a saturated hydrocarbon chain and an unsaturated hydrocarbon chain. 
     
     
         7 . The lipid construct of  claim 7 , wherein the backbone comprises glycerol. 
     
     
         8 . The lipid construct of  claim 1 , wherein the distal moiety of the extended amphipathic lipid comprises at least one member selected from the group consisting of biotin, a biotin derivative, iminobiotin, an iminobiotin derivative, biocytin, a biocytin derivative, iminobiocytin, an iminobiocytin derivative and a hepatocyte specific molecule that binds to a receptor in a hepatocyte. 
     
     
         9 . The lipid-construct of  claim 1  where the extended amphipathic lipid is selected from the group consisting of N-hydroxysuccinimide (NHS) biotin; sulfo-NHS-biotin; N-hydroxysuccinimide long chain biotin; sulfo-N-hydroxysuccinimide long chain biotin; D-biotin; biocytin; sulfo-N-hydroxysuccinimide-S-S-biotin; biotin-BMCC; biotin-HPDP; iodoacetyl-LC-biotin; biotin-hydrazide; biotin-LC-hydrazide; biocytin hydrazide; biotin cadaverine; carboxybiotin; photobiotin; ρ-aminobenzoyl biocytin trifluoroacetate; ρ-diazobenzoyl biocytin; biotin DHPE; biotin-X-DHPE; 12-((biotinyl)amino)dodecanoic acid; 12-((biotinyl)amino)dodecanoic acid succinimidyl ester; S-biotinyl homocysteine; biocytin-X; biocytin x-hydrazide; biotmethylenediamine; biotin-XL; biotin-X-ethylenediamine; biotin-XX hydrazide; biotin-XX-SE; biotin-XX, SSE; biotin-X-cadaverine; α-(t-BOC)biocytin; N-(biotinyl)-N′-(iodoacetyl)ethylenediamine; DNP-X-biocytin-X-SE; biotin-X-hydrazide; norbiotinamine hydrochloride; 3-(N-maleimidylpropionyl)biocytin; ARP; biotin-1-sulfoxide; biotin methyl ester; biotin-maleimide; biotin-poly(ethyleneglycol)amine; (+) biotin 4-amidobenzoic acid sodium salt; Biotin 2-N-acetylamino-2-deoxy-β-D-glucopyranoside; Biotin-α-D-N-acetylneuraminide; Biotin-α-L-fucoside; Biotin lacto-N-bioside; Biotin-Lewis-A trisaccharide; Biotin-Lewis-Y tetrasaccharide; Biotin-α-D-mannopyranoside; biotin 6-O-phospho-α-D-mannopyranoside; and polychromium-poly(bis)-N-[2,6-(diisopropylphenyl) carbamoyl methylimino] diacetic acid. 
     
     
         10 . The lipid construct of  claim 1 , wherein the medial moiety of the extended amphipathic lipid comprises a thio-acetyl triglycine polymer or a derivative thereof, wherein the extended amphipathic lipid molecule extends outward from the surface of the lipid construct. 
     
     
         11 . The lipid construct of  claim 2 , further comprising at least one insulin associated with a water insoluble target molecule complex, wherein the complex comprises a plurality of linked individual units, wherein the individual units comprise:
 a. a bridging component selected from the group consisting of a transition element, an inner transition element, a neighbor element of the transition element and a mixture of any of the foregoing elements, and   b. a complexing component,   
       provided that when the transition element is chromium, a chromium target molecule complex is formed. 
     
     
         12 . The lipid construct of  claim 11 , further comprising at least one insulin that is not associated with the target molecule complex. 
     
     
         13 . The lipid construct of  claim 11 , wherein the bridging component is chromium. 
     
     
         14 . The lipid construct of  claim 11 , wherein the complexing component comprises poly(bis)-[(N-(2,6-diisopropylphenyl)carbamoyl methyl) iminodiacetic acid]. 
     
     
         15 . The lipid construct of  claim 1 , wherein the distal component of the extended amphipathic lipid comprises a non-polar derivatized benzene ring or a heterobicyclic ring structure. 
     
     
         16 . The lipid construct of  claim 1 , wherein the construct comprises a positive charge, a negative charge or combinations thereof. 
     
     
         17 . The lipid construct of  claim 1 , wherein the extended amphipathic lipid comprises at least one carbonyl moiety positioned at a distance about 13.5 angstroms or less from the terminal end of the distal moiety. 
     
     
         18 . The lipid construct of  claim 1 , wherein the extended amphipathic lipid comprises at least one carbamoyl moiety comprising a secondary amine. 
     
     
         19 . The lipid construct of  claim 1 , wherein the extended amphipathic lipid comprises charged chromium in the medial position. 
     
     
         20 . The lipid construct of  claim 1 , further comprising cellulose acetate hydrogen phthalate. 
     
     
         21 . The lipid construct of  claim 2 , further comprising at least one charged organic molecule bound to the insulin. 
     
     
         22 . The lipid construct of  claim 2 , wherein the charged organic molecule is selected from the group consisting of protamines, derivatives of polylysine, highly basic amino acid polymers, poly (arg-pro-thr) n  in a mole ratio of 1:1:1, poly (DL-Ala-poly-L-lys) n  in a mole ratio of 6:1, histones, sugar polymers that contain a positive charge contributed by a primary amino group, polynucleotides with primary amino groups, carboxylated polymers and polymeric amino acids, fragments of proteins that contain large amounts of amino acid residues with carboxyl (COO − ) or sulthydral (S − ) functional groups, derivative of proteins with negatively charged terminal acidic carboxyl groups, acidic polymers, sugar polymers containing negatively charged carboxyl groups, a derivative thereof, and any combination of the aforementioned compounds. 
     
     
         23 . A method of manufacturing a lipid construct comprising an amphipathic lipid and an extended amphipathic lipid, wherein the extended amphipathic lipid comprises proximal, medial and distal moieties, wherein the proximal moiety connects the extended amphipathic lipid to the construct, the distal moiety targets the construct to a receptor displayed by a hepatocyte, and the medial moiety connects the proximal and distal moieties, the method comprising:
 a. creating a mixture comprising the amphipathic lipid and an extended amphipathic lipid; and   b. forming a suspension of the lipid construct in aqueous media.   
     
     
         24 . A method of manufacturing the lipid construct comprising an insulin, an amphipathic lipid and an extended amphipathic lipid, wherein the extended amphipathic lipid comprises proximal, medial and distal moieties, wherein the proximal moiety connects the extended amphipathic lipid to the construct, the distal moiety targets the construct to a receptor displayed by a hepatocyte, and the medial moiety connects the proximal and distal moieties, the method comprising:
 a. creating a mixture comprising the amphipathic lipid and an extended amphipathic lipid;   b. forming a suspension of the lipid construct in aqueous media; and   c. loading the insulin into the lipid construct.   
     
     
         25 . The method of  claim 24 , wherein the step of loading the insulin into the lipid construct comprises equilibrium loading and non-equilibrium loading. 
     
     
         26 . The method of  claim 24 , wherein the step of loading the insulin into the lipid construct comprises adding a solution containing free insulin to a mixture of the lipid construct in aqueous media and allowing the insulin to remain in contact with the mixture until equilibrium is reached. 
     
     
         27 . The method of  claim 24 , further comprising the step of
 d. terminally loading the insulin into the lipid construct after the mixture reaches equilibrium, wherein the solution containing free insulin is removed from the construct, further wherein the construct comprises insulin.   
     
     
         28 . The method of  claim 27 , further comprising the step of:
 e. removing the solution containing free insulin from the lipid construct containing insulin associated with the construct by a process selected from the group consisting of a rapid filtration procedure, centrifugation, filter centrifugation, and chromatography using an ion-exchange resin or streptavidin agarose affinity-resin gel having affinity for biotin, iminobiotin or derivates thereof.   
     
     
         29 . The method of  claim 24 , further comprising the step of:
 f. adding a chromium complex comprising a plurality of linked individual units to the lipid construct.   
     
     
         30 . The method of  claim 24 , further comprising the step of:
 g. adding cellulose acetate hydrogen phthalate to the lipid construct.   
     
     
         31 . The method of  claim 24  further comprising the step of:
 h. reclaiming from the process at least one material selected from the group consisting of insulin, ion-exchange resin and streptavidin agarose affinity-gel. 
 
     
     
         32 . The method of  claim 24 , wherein the step of loading the insulin into the lipid construct comprises the step of adding at least one charged organic molecule to the insulin before the insulin is loaded into the lipid construct. 
     
     
         33 . A method of increasing the bioavailability of at least one insulin in a patient comprising:
 a. combining at least one insulin with a lipid construct, wherein the lipid construct comprises a plurality of non-covalent multi-dentate binding sites; and   b. administering the construct containing insulin to the patient.   
     
     
         34 . The method of  claim 33 , further comprising the step of modulating the isoelectric point of at least one active ingredient. 
     
     
         35 . The method of  claim 33 , wherein the insulin is selected from the group consisting of insulin lispro, insulin aspart, regular insulin, insulin glargine, insulin zinc, human insulin zinc extended, isophane insulin, human buffered regular insulin, insulin glulisine, recombinant human regular insulin, recombinant human insulin isophane, premixed combinations of any of the aforementioned insulins, a derivative thereof, and a combination of any of the aforementioned insulins. 
     
     
         36 . The method of  claim 33 , wherein the lipid construct comprises insulin, 1,2-distearoyl-sn-glycero-3-phophocholine, cholesterol, dicetyl phosphate, 1,2-dipalmitoyl-sn-glycero-[3-phospho-rac-(1-glycerol)], 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl) or derivatives, and a hepatocyte receptor binding molecule. 
     
     
         37 . The method of  claim 33 , further comprising the step of adding at least one charged organic molecule to the insulin before the insulin is combined with the lipid construct. 
     
     
         38 . A method of forming a time-release composition that provides increased bio-distribution of insulin in a host comprising:
 a. removing a lipid construct from a bulk phase media by binding the construct through lipids comprising iminobiotin or an iminobiotin derivative to streptavidin agarose affinity-gel at pH 9.5 or greater;   b. separating the construct from the bulk phase media; and   c. releasing the construct from the affinity-gel by adjusting the pH of an aqueous mixture of the affinity gel to pH 4.5, wherein, the released construct contains insoluble insulin;   wherein upon administration of the construct to a warm-blooded host the insulin is resolubilized under the physiological pH conditions in the host.   
     
     
         39 . A method of treating a patient afflicted with diabetes comprising administering to the patient an effective amount of a lipid construct comprising insulin associated with the construct. 
     
     
         40 . The method of  claim 39 , wherein the insulin is selected from the group consisting of insulin lispro, insulin aspart, regular insulin, insulin glargine, insulin zinc, human insulin zinc extended, isophane insulin, human buffered regular insulin, insulin glulisine, recombinant human regular insulin, recombinant human insulin isophane, premixed combinations of any of the aforementioned insulins, a derivative thereof, and a combination of any of the aforementioned insulins. 
     
     
         41 . The method of  claim 39 , wherein the lipid construct further comprises a target molecule complex, wherein the complex comprises a plurality of linked individual units, further wherein the linked individual units comprise:
 a. a bridging component selected from the group comprising a transition element, an inner transition element, a neighbor element of the transition element and a mixture of any of the foregoing elements; and   b. a complexing component;   provided that when the transition element is chromium, a chromium target molecule complex is formed.   
     
     
         42 . The method of  claim 39 , wherein the lipid construct further comprises insulin not associated with the target molecule complex. 
     
     
         43 . The method of  claim 39 , wherein the administration is oral or subcutaneous. 
     
     
         44 . The method of  claim 39 , wherein the insulin associated with the construct comprises at least one charged organic molecule bound to the insulin. 
     
     
         45 . A method for increasing the delivery of insulin to hepatocytes in the liver of a patient afflicted with diabetes by administering to the patient a lipid construct comprising insulin, an amphipathic lipid, and an extended lipid, wherein the extended lipid comprises a moiety that binds to hepatocyte receptors, wherein the lipid construct is present in a plurality of sizes. 
     
     
         46 . The method of  claim 45 , wherein the insulin is selected from the group consisting of insulin lispro, insulin aspart, regular insulin, insulin glargine, insulin zinc, human insulin zinc extended, isophane insulin, human buffered regular insulin, insulin glulisine, recombinant human regular insulin, recombinant human insulin isophane, premixed combinations of any of the aforementioned insulins, a derivative thereof, and a combination of any of the aforementioned insulins. 
     
     
         47 . The method of  claim 45 , further comprising protecting the insulin within the lipid construct from hydrolytic degradation by providing a three-dimensional structural array of lipid molecules so as to prevent access to the insulin by hydrolytic enzymes. 
     
     
         48 . The method of  claim 45 , further comprising adding cellulose acetate hydrogen phthalate to the lipid construct to react with individual lipid molecules. 
     
     
         49 . The method of  claim 45 , further comprising producing an insolubilized dosage form of insulin within the lipid construct. 
     
     
         50 . A kit for use in treating a mammal inflicted with diabetes, the kit comprising a lipid construct, a physiological buffer solution, an applicator, and an instructional material for the use thereof, wherein the lipid construct comprises an amphipathic lipid and an extended amphipathic lipid, wherein the extended amphipathic lipid comprises proximal, medial and distal moieties, wherein the proximal moiety connects the extended amphipathic lipid to the construct, the distal moiety targets the construct to a receptor displayed by a hepatocyte, and the medial moiety connects the proximal and distal moieties. 
     
     
         51 . The kit of  claim 50 , further comprising at least one insulin. 
     
     
         52 . A hepatocyte-targeting composition comprising:
 a. at least one free insulin; and   b. at least one insulin associated with a water-insoluble target molecule complex;   wherein the target molecule complex is comprised of a combination of:   a. multiple linked individual units, the individual units comprising:
 i. at least one bridging component selected from the group consisting of a transition element, an inner transition element, and a neighbor element of the transition element; and 
 ii. a complexing component; and 
   b. a lipid construct matrix comprising at least one lipid component;   provided that when the transition element is chromium, a chromium target molecule complex is created;   further wherein the target molecule complex comprises a negative charge.   
     
     
         53 . The hepatocyte-targeting composition of  claim 52 , wherein the insulin is selected from the group consisting of insulin lispro, insulin aspart, regular insulin, insulin glargine, insulin zinc, human insulin zinc extended, isophane insulin, human buffered regular insulin, insulin glulisine, recombinant human regular insulin, recombinant human insulin isophane, premixed combinations of any of the aforementioned insulins, a derivative thereof, and a combination of any of the aforementioned insulins. 
     
     
         54 . The hepatocyte targeted composition of  claim 52 , wherein the insulin comprises an insulin-like moiety, including a fragments of an insulin molecules, that has a biological activity of insulin. 
     
     
         55 . The hepatocyte-targeting composition of  claim 52 , wherein the lipid component comprises at least one lipid selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2-dimyristoyl-sn-glycero-3-phosphocholine, cholesterol, cholesterol oleate, dicetylphosphate, 1,2-distearoyl-sn-glycero-3-phosphate, 1,2-dipalmitoyl-sn-glycero-3-phosphate, and 1,2-dimyristoyl-sn-glycero-3-phosphate. 
     
     
         56 . The hepatocyte-targeting composition of  claim 52 , wherein the lipid component comprises at least one lipid selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine, cholesterol, and dicetyl phosphate. 
     
     
         57 . The hepatocyte-targeting composition of  claim 52 , wherein the lipid component comprises a mixture of 1,2-distearoyl-sn-glycero-3-phosphocholine, cholesterol and dicetyl phosphate. 
     
     
         58 . The hepatocyte-targeting composition of  claim 52 , wherein the bridging component is chromium. 
     
     
         59 . The hepatocyte-targeting composition of  claim 52 , wherein the complexing component comprises at least one member selected from the group consisting of:
 N-(2,6-diisopropylphenylcarbamoylmethyl) iminodiacetic acid;   N-(2,6-diethylphenylcarbamoylmethyl) iminodiacetic acid;   N-(2,6-dimethylphenylcarbamoylmethyl) iminodiacetic acid;   N-(4-isopropylphenylcarbamoylmethyl) iminodiacetic acid;   N-(4-butylphenylcarbamoylmethyl) iminodiacetic acid;   N-(2,3-dimethylphenylcarbamoylmethyl) iminodiacetic acid;   N-(2,4-dimethylphenylcarbamoylmethyl) iminodiacetic acid;   N-(2,5-dimethylphenylcarbamoylmethyl) iminodiacetic acid;   N-(3,4-dimethylphenylcarbamoylmethyl) iminodiacetic acid;   N-(3,5-dimethylphenylcarbamoylmethyl) iminodiacetic acid;   N-(3-butylphenylcarbamoylmethyl) iminodiacetic acid;   N-(2-butylphenylcarbamoylmethyl) iminodiacetic acid;   N-(4-tertiary butylphenylcarbamoylmethyl) iminodiacetic acid;   N-(3-butoxyphenylcarbamoylmethyl) iminodiacetic acid;   N-(2-hexyloxyphenylcarbamoylmethyl) iminodiacetic acid;   N-(4-hexyloxyphenylcarbamoylmethyl) iminodiacetic acid; aminopyrrol iminodiacetic acid;   N-(3-bromo-2,4,6-trimethylphenylcarbamoylmethyl) iminodiacetic acid; benzimidazole methyl iminodiacetic acid;   N-(3-cyano-4,5-dimethyl-2-pyrrylcarbamoylmethyl) iminodiacetic acid;   N-(3-cyano-4-methyl-5-benzyl-2-pyrrylcarbamoylmethyl) iminodiacetic acid; and   N-(3-cyano-4-methyl-2-pyrrylcarbamoylmethyl) iminodiacetic acid.   
     
     
         60 . The hepatocyte-targeting composition of  claim 52 , wherein the complexing component comprises poly(bis)[N-(2,6-diisopropylphenylcarbamoylmethyl) iminodiacetic acid]. 
     
     
         61 . A method of manufacturing a hepatocyte-targeting composition of  claim 52 , comprising:
 creating a target molecule complex, wherein the complex comprises multiple linked individual units and a lipid construct matrix;   forming a suspension of the target molecule complex in buffer; and   combining the insulin and the target molecule complex.   
     
     
         62 . A method of manufacturing a hepatocyte-targeting composition of  claim 52 , comprising:
 creating a target molecule complex, wherein the complex comprises multiple linked individual units and a lipid construct matrix;   forming a suspension of the target molecule complex in aqueous media;   adjusting the pH of the aqueous suspension to approximately pH 5.3;   adjusting the pH of the glargine insulin to approximately 4.8; and   combining the glargine insulin and the target molecule complex,   wherein the insulin is glargine insulin.   
     
     
         63 . A method of manufacturing a hepatocyte-targeting composition of  claim 52 , comprising:
 creating a target molecule complex, wherein the complex comprises multiple linked individual units and a lipid construct matrix;   forming a suspension of the target molecule complex in aqueous media;   adjusting the pH of the aqueous suspension to approximately pH 5.3;   adjusting the pH of the glargine insulin to approximately 4.8; and   combining the glargine insulin, the non-glargine insulin and the target molecule complex,   wherein the insulin comprises glargine insulin and at least one non-glargine insulin.   
     
     
         64 . A method of treating a patient for Type I or Type II diabetes comprising administering to the patient an effective amount of a hepatocyte-targeting composition of  claim 52 . 
     
     
         65 . The method of treating a patient according to  claim 64 , wherein the route of administration is selected from the group consisting of oral, parenteral, subcutaneous, pulmonary and buccal. 
     
     
         66 . The method of treating a patient according to  claim 64 , wherein the route of administration is oral or subcutaneous. 
     
     
         67 . A method of treating a patient for Type I or Type II diabetes comprising administering to the patient an effective amount of a hepatocyte targeted composition of  claim 52 , wherein insulin comprises glargine insulin and at least one non-glargine insulin, further wherein the non-glargine insulin is selected from the group consisting of insulin lispro, insulin aspart, regular insulin, insulin glargine, insulin zinc, human insulin zinc extended, isophane insulin, human buffered regular insulin, insulin glulisine, recombinant human regular insulin, recombinant human insulin isophane, premixed combinations of any of the aforementioned insulins, a derivative thereof, and a combination of any of the aforementioned insulins. 
     
     
         68 . The method of  claim 67 , wherein the non-glargine insulin comprises insulin-like moieties, including fragments of insulin molecules, that have biological activity of insulins. 
     
     
         69 . A method of treating a patient for Type I or Type II diabetes comprising administering to the patient an effective amount of a hepatocyte-targeting composition of  claim 52 . 
     
     
         70 . The method of treating a patient according to  claim 69 , wherein the route of administration is selected from the group consisting of oral, parenteral, subcutaneous, pulmonary and buccal. 
     
     
         71 . The method of treating a patient according to  claim 69 , wherein the route of administration is oral or subcutaneous. 
     
     
         72 . A method of treating a patient for Type I or Type II diabetes comprising administering to the patient an effective amount of a hepatocyte targeted composition of  claim 51 , wherein insulin comprises recombinant human insulin isophane and at least one insulin that is not recombinant human insulin isophane. 
     
     
         73 . The method of  claim 72 , wherein the at least one insulin that is not recombinant human insulin isophane comprises insulin-like moieties, including fragments of insulin molecules, that have biological activity of insulins. 
     
     
         74 . A kit for use in treating Type I or Type II diabetes in a mammal, the kit comprising a physiological buffered solution, an applicator, instructional material for the use thereof, and a water insoluble target molecule complex, wherein the complex comprises multiple linked individual units and a lipid construct matrix containing a negative charge, the multiple linked individual units comprising: a bridging component selected from the group consisting of a transition element, an inner transition element, a neighbor element of the transition element and a mixture of any of the foregoing elements, and a complexing component, provided that when the transition element is chromium, a chromium target molecule complex is created, wherein the multiple linked individual units are combined with the lipid construct matrix. 
     
     
         75 . The kit of  claim 74  further comprising at least one insulin, wherein the insulin is associated with the target molecule complex, wherein the complex comprises a charge.