Abstract: A novel organic anion transporter gene participating in organic anion transport in the placenta; and an organic anion transporter which is a polypeptide encoded by the gene. A placental organic anion transporter OAT4, more particularly, a placental organic anion transporter OAT4 having the amino acid sequence represented by SEQ ID NO:2 or an amino acid sequence derived therefrom by deletion, substitution or addition of a part of the amino acids thereof. A nucleic acid (preferably DNA) having a base sequence encoding the placental organic anion transporter OAT4 or a base sequence hybridizable therewith under stringent conditions.

Abstract: Disclosed is a method for producing seeding microcrystals for the production of human insulin, the microcrystals being free of non-human pancreatic insulin, the method comprising providing an unseeded suspension of human insulin, the suspension being free of non-human pancreatic insulin, and homogenizing the insulin suspension under pressure to result in human insulin microcrystals suitable for use as seeding microcrystals for the production of zinc insulin products. The method of homogenization under pressure may also be used for the production of seeding mnicrocrystals for other peptides and proteins, in particular pharmaceutical peptides or proteins such as insulin, GLP-1, glucagon and growth hormones.

Abstract: This invention relates generally to a multimeric polypeptide complex/antigen which may be utilized in detecting or diagnosing diseases of the breast such as breast cancer. Furthermore, the invention relates to methods and kits, for example, which utilize this antigen or an antibody thereto. The complex itself comprises at least one BU101 polypeptide and at least one Mammaglobin polypeptide. In addition, the complex may comprise at least one polypeptide having at least 20% identity with the amino acid sequence of the BU101 polypeptide, the amino acid sequence of the Mammaglobin polypeptide, and fragments thereof.

Abstract: The present invention provides angiogenic vasculature homing molecules that bind to NG2/HM proteoglycan, including, for example, a peptide comprising the amino acid sequence TAASGVRSMH (SEQ ID NO:1) or LTLRWVGLMS (SEQ ID NO:2). The invention also provides conjugates comprising an angiogenic vasculature homing molecule linked to a moiety such as a drug, a cytotoxic agent, a chemotherapeutic agent, or a detectable agent. The invention additionally provides a method of targeting angiogenic vasculature in a tumor in vivo by contacting the angiogenic vasculature with an angiogenic vasculature homing molecule that selectively homes to a NG2/HM proteoglycan, wherein the angiogenic vasculature homing molecule is not an antibody.

Abstract: The present invention relates to a vector for expression of a heterologous protein by a Gram negative bacteria, wherein the vector includes a nucleic acid such as DNA encoding the following: an origin of replication region; optionally and preferably a selection marker; a promoter; an initiation region such as translation initiation region and/or a ribosome binding site, at least one restriction site for insertion of heterologous nucleic acid, e.g. DNA, encoding the heterologous protein, and a transcription terminator. The inventive vector may contain DNA encoding the heterologous protein, e.g., pro-insulin such as pro-insulin with a His tag.

Abstract: The present invention relates to an improved process for obtaining a precursor of an insulin or insulin derivative having correctly bonded cystine bridges in the presence of cysteine or cysteine hydrochloride and chaotropic auxiliary.

Abstract: The invention provides methods and compositions useful for making synthetic peptide conjugates. In one embodiment, the invention provides compositions comprising the structure: wherein R is selected from lower substituted or unsubstituted alkyl, O, NH and S and P is an amine protection group. In more particular embodiments, the compositions comprise &agr;-amine protected 4,5-dehydroleucine or &agr;-amine protected (2S)-aminolevulinic acid and/or P is F-moc. These compounds may be incorporated into peptides, for example, peptides comprising a substituted or unsubstituted (2S)-aminolevulinic acid residue, such as (2S)-aminolevulinic acid residue is substituted with an O- or N-linked glycoconjugate, or a detectable label.

Abstract: The current invention provides antioxidative peptide fractions. The invention includes antioxidative peptides of casein having sequences of SEQ ID NOS:1-5. The invention includes methods for making or generating the peptides, isolated nucleic acids encoding the peptides, and expression. vectors comprising these nucleic acids. The invention includes food additives for preventing oxidation in situ. The invention includes stabilized products with improved storage for products ingredients that are subject to oxidation. Finally, the invention includes methods for isolating peptide fractions having antioxidative activity from protein samples.

Abstract: The present invention relates to novel human secreted proteins and isolated nucleic acids containing the coding regions of the genes encoding such proteins. Also provided are vectors, host cells, antibodies, and recombinant methods for producing human secreted proteins. The invention further relates to diagnostic and therapeutic methods useful for diagnosing and treating disorders related to these novel human secreted proteins.

Abstract: A peptide of the formula (I) (wherein each symbol is as defined in the description.) or an acid addition salt thereof. A compound of the formula (I) is a specific substrate for human pepsin II, so it can be used for assaying human pepsin II or human pepsinogen II and it is useful for diagnosis of gastric diseases such as gastric cancer and gastric ulcer.

Abstract: The present invention relates to insoluble compositions containing acylated proteins selected from the group consisting of acylated insulin, acylated insulin analog, and acylated proinsulin, and formulations thereof. The formulations are suitable for parenteral delivery or other means of delivery, to a patient for extended control of blood glucose levels. More particularly, the present invention relates to compositions comprised of an acylated protein complexed with zinc, protamine, and a phenolic compound such that the resulting microcrystal is analogous to the neutral protamine Hagedorn (NPH) insulin crystal form. Surprisingly, it has been discovered that compositions of such acylated proteins have therapeutically superior subcutaneous release pharmacokinetics, and more extended and flatter glucodynamics, than presently available commercial preparations of NPH insulin.

Abstract: The present invention relates to zinc free insulin crystals having a diameter below 10 &mgr;m and to therapeutic powder formulations suitable for pulmonary administration containing such insulin crystals. The crystals of the present invention exhibit a better stability profile than powders of essentially the same composition prepared by spray drying, freeze-drying, vacuum drying and open drying. The therapeutic powder formulations elucidate better flowing properties than corresponding amorphous powder formulations.

Abstract: This invention relates to certain peptides and linear and cyclic analogs thereof that bind to the mu (morphine) opiate receptor with higher affinity, selectivity and potency than currently available peptides. This invention also relates to pharmaceutical preparations containing an effective amount of the peptides or salts thereof, and methods for providing analgesia, relief from gastrointestinal disorders such as diarrhea, and therapy for drug dependence containing a pharmaceutically effective amount of the peptides.

Abstract: The present invention relates to a vector for expression of a heterologous protein by a Gram negative bacteria, wherein the vector includes a nucleic acid such as DNA encoding the following: an origin of replication region; optionally and preferably a selection marker; a promoter; an initiation region such as translation initiation region and/or a ribosome binding site, at least one restriction site for insertion of heterologous nucleic acid, e.g. DNA, encoding the heterologous protein, and a transcription terminator. The inventive vector may contain DNA encoding the heterologous protein, e.g., pro-insulin such as pro-insulin with a His tag.

Abstract: A method for culturing a recombinant host cell comprising: determining a polypeptide factor for a polypeptide factor-dependent host cell; transforming said host cell with nucleic acid encoding said polypeptide factor; transforming the host cell with nucleic acid encoding a desired protein; and, culturing the transformed host cells in a medium lacking the polypeptide factor.

Abstract: The invention concerns a process of chromatographically separating glycosylated proteins from non-glycosylated proteins by subjecting a solution comprising glycosylated and non-glycosylated proteins to chromatography using a Ca++ containing eluant. By using this process a fraction comprising non-glycosylated proteins substantially free from glycosylated proteins is obtained. The process may be applied to the separation of proteins used in the medical industry, such as insulin.

Abstract: An aqueous insulin composition with improved chemical and physical stability comprising human insulin or an analogue or derivative thereof, a buffer selected from glycylglycine, citrate or TRIS, in particular glycylglycine, and metal ions, in particular calcium or magnesium ions.

Abstract: Divalent cation crystals of human growth factor (hGH) or derivatives thereof, and pharmaceutical preparations comprising divalent cation crystals of hGH. In specific embodiments, the divalent cation is Zn++ and the molar ration Zn++ and hGH is about 0.2 to about 10.

Abstract: Low doses of stroma-free diaspirin cross-linked hemoglobin are administered to patients undergoing hemodialysis, to achieve hemostabilization and avoid hypotensive episodes in susceptible patients. Hemoglobin therapy when implemented prophylactically in hemodialysis also partially obviates the need for further interventions to control circulatory system instability.

Abstract: A process for isolating insulin which entails performing high-pressure liquid chromatography with pressure-stable acidic cation exchange material under a pressure of from 1.1 MPa to 40 MPa.

Abstract: A receptor for an insulin-like polypeptide is purified from yeast membranes. This "insulin receptor-like protein" has a structure analogous to that of the mammalian insulin receptor. The insulin receptor-like protein of Saccharomyces cerevisiae is a heterotetrameric glycoprotein. The protein has two polypeptide types, a first polypeptide which binds insulin and has an apparent molecular weight of 135,000 to 145,000 daltons and a second polypeptide which has an apparent molecular weight of 90,000 to 95,000 daltons and is phosphorylated on tyrosine in response to binding of insulin by said first polypeptide. The first and second polypeptides are joined by disulfide linkage. The protein requires a divalent metal ion for tyrosine autophosphorylation in response to binding of insulin. The yeast insulin receptor-like protein binds human insulin with a dissociation constant of K.sub.d =8.times.10.sup.-10 M and binds human insulin-like growth factor 1 with a K.sub.d =4.times.10.sup.-10 M.

Abstract: The present invention relates to a process for preparing human proinsulin which is represented as a following chemical formula(I): ##STR1## wherein, R is an amino acid residue or a peptide which is degradable enzymatically or chemically; and, X is a linkage of an amino group of A-1 in insulin A chain and a carboxyl group of B-30 in insulin B chain which can be separated from the A chain or the B chain enzymatically or chemically, provided that a region from A-1 to A-21 is the insulin A chain and a region from B-1 to B-30 is the insulin B chain. In accordance with the present invention, human recombinant insulin precursor can be simply manufactured with a good reproducibility, since dissolution, sulfonation, concentration, desalting and purification are remarkably simplified, while increasing the yield of refolding reaction.

Abstract: A new use of hydrophobic zeolites, viz. for removing preservatives from (poly)peptide solutions, e.g. solutions of pharmaceutical preparations. The use is of particular interest in connection with protein solutions, especially insulin solutions. The invention further relates to an injection syringe for such solutions, in which syringe a zeolite of the type at issue is placed for contact with the solution to remove preservatives therefrom.

Abstract: A method of providing zinc containing crystals of a protein derivative which has a lysine residue which carries a lipophilic substituent on the .epsilon.-amino group, said method comprising providing a solution of the protein derivative in an alkaline buffer, which further contains a zinc salt, adjusting the pH value of the solution to a value between 7 and 10, and isolating the crystals formed.

Abstract: The present invention relates to insulin crystals comprising ASP.sup.B28 and protamine, and pharmaceutical preparations containing same. The crystals and preparations exhibit rapid onset and prolonged activity when administered in vivo.

Abstract: The present invention relates to insulin crystals comprising ASP.sup.B28 and protamine, and pharmaceutical preparations containing same, The crystals and preparations exhibit rapid onset and prolonged activity when administered in vivo.

Abstract: Insulin formulations containing ligands for the insulin hexamer which bind several orders of magnitude more tightly to the hexamer than chlorine ion or acetate ion. These ligands are aliphatic and aromatic carboxylates having a dissociation constant (K.sub.D) of less than about 5 mM, and preferably less than about 1 mM. The increased tightness of binding conveys additional stability to the insulin hexamers, improving their usefulness in slow release and fast acting formulations (for example, for the treatment of diabetics).

Abstract: A method of producing chemically stable and biologically active growth hormone crystals and processes for production of pharmaceutical preparations containing these growth hormone crystals.

Abstract: A method for recovering an acylated protein as a powder, especially in cases where said acylated protein is one that resists isolation by isoelectric precipitation from aqueous solutions of the protein, said method comprising in combination adjusting said aqueous solution to near the isoelectric pH of the protein and providing a suitable alcohol concentration to cause precipitation of the protein in the form of filterable particles at said pH.

Abstract: A process for obtaining insulin having correctly linked cystine bridges from a corresponding proinsulin amino acid chain in the presence of mercaptan, chaotropic auxiliaries and at a pH of 10 to 11 is described, as well as the direct conversion of the proinsulin to insulin with the aid of enzymes and direct isolation of the insulin from the reaction mixture with the aid of hydrophobic adsorber resins.

Abstract: The present invention discloses various parenteral pharmaceutical formulations, which comprise: a monomeric insulin analog, zinc, protamine, and phenolic derivative. The analog formulations provide a prolonged duration of action. A process for preparing insulin analog-protamine formulations is also described.

Abstract: A method for processing a fatty acid-acylated protein, especially N-palmitoyl Lys.sup.B29 human insulin, to reduce the incidence of gelation by conducting such processing in the presence of a citrate buffering agent as the primary buffer which method allows such processing to be conducted at higher protein concentrations and with less temperature control than would otherwise be possible in the absence of the citrate buffer.

Abstract: A process is described for obtaining insulin, which is virtually free from proteases and/or insulin acetylated at position A9, by chromatography in aqueous, buffered solvents which contain water-miscible organic solvents, on lipophilically modified silica gel, wherein the dissolving buffer contains acetone or acetonitrile.

Abstract: Analogs of human insulin containing an aspartic acid at position 1 of the B chain (Asp.sup.B1), and optionally, having a gln modification at position 13 (Gln.sup.B13), display modified physico-chemical and pharmacokinetic properties which enable them to be long acting. These analogs are useful in the treatment of hyperglycemia because they are soluble and display an increased tendency to self-associate.

Abstract: The present invention discloses various parenteral pharmaceutical formulations, which comprise a mixture of a monomeric insulin analog and insulin-NPH. The analog formulations provide a rapid onset and a prolonged duration of action.

Abstract: The present invention relates to insulin crystals comprising ASP.sup.B28 and protamine, and pharmaceutical preparations containing same. The crystals and preparations exhibit rapid onset and prolonged activity when administered in vivo.

Abstract: This specification describes a process for converting a human insulin precursor to human insulin, which comprises treating such human insulin precursor with trypsin and carboxypeptidase B in an aqueous medium containing per mole of human insulin precursor from about 0.1 to about 10 moles of one or more metal ions of those metals having Atomic Numbers 21 to 34, 39 to 52, 57 to 84, and 89 to 92.

Abstract: A process for phosphorylating a peptide comprises reacting an aqueous solution of the peptide with an effective amount of phosphorous oxychloride under conditions favoring phosphorylation of the peptide. In a specific embodiment, the peptide is insulin. A phosphorylated insulin is used in the treatment of diabetes mellitus wherein the phosphorylated insulin is produced or purified with an effective amount of phosphorus oxychloride to have substantially reduced iso-electric points and to have the property of reducing hyperglycemia without substantially inducing hypoglycemia. In a specific embodiment of the invention, the method comprises administering to a human being an effective therapeutic amount of a phosphorylated insulin essentially free of unmodified insulin and having substantially reduced iso-electric points whereby hyperglycemia is reduced substantially without inducing hypoglycemia.

Abstract: Novel insulin compounds having a desirably protracted insulin action and/or antigenicity are provided.The novel insulin compounds are represented by the formula II: ##STR1## wherein E individually represents Glu or a neutral amino acid residue which can be coded for by nucleotide sequences, N represents an amino acid residue which can be coded for by nucleotide sequences,T represents Thr or Arg,X represents Thr, Ser, Ala or OH, andY represents OR or NR.sup.1 R.sup.2, where R, R.sup.1 and R.sup.2 individually represents hydrogen or lower alkyl, but is not present when X represents OH.

Abstract: This disclosure relates to new compounds and to a process for the controlled conversion of such compounds to produce a Protein that comprises an amino acid sequence which is useful as a precursor to human insulin or to a modified human insulin, the amino terminus portion of which sequence does not define a cathepsin C dipeptide removal stop point. The compounds have the formula Met-Y-Protein in which Y is Tyr or Arg.

Abstract: In a process for the production of a soluble native protein, such as immunoglobulin or methionine-prochymosin, in which an insoluble form of the protein is produced by a host organism transformed with a vector including a gene coding for the protein, the insoluble form of the protein is reversibly denatured in an alkaline aqueous solution at a pH selected to promote dissociation of a group or groups of the protein involved in maintaining the conformation of the protein, and the protein is subsequently allowed to renature by reducing the pH of the solution below a pH effective to denature the protein to produce the soluble native form of the protein. The pH of the alkaline aqueous is suitably in the range 9.0 to 11.5.

Abstract: The production of recombinant chymosin is disclosed in which an insoluble form of chymosin precursor is produced by a bacterial host cell transformed by a vector including a coding sequence for said precursor. Solubilization of said insoluble form of chymosin precursor is accomplished using urea at a concentration of at least 7M or guanidine hydrochloride at a concentration of at least 6M prior to cleaving said precursor to form chymosin. Said solubilization preferably additionally involves the denaturation of said precursor in an alkaline aqueous solution, e.g., at a pH between a 9 and 11.

Abstract: A method of removing a valuable protein from a complex solution and recovering the valuable protein in purified form consists of adding a silica gel sorbant having a pore size approximately the molecular size of the protein to a solution containing the protein, allowing the protein to be sorbed onto the sorbant, separating the sorbant from the solution and then recovering the protein from the sorbant.

Abstract: A process for the purification of insulins and insulin derivatives by chromatography in aqueous, buffered solvents which contain water-miscible organic solvents, on lipophilically modified silica gel, is described, wherein the aqueous, buffered solvents contain zwitterions, or the pH of the solvent mixture is in the vicinity of the isoelectric point of the insulin or insulin derivative to be purified and zwitterions are present.

Abstract: Phosphorylated insulins which can be prepared from chemically extracted pharmacological insulins by gentle treatment with phosphorus oxychloride, have been shown to have reduced bioactivity by mouse convulsion assay, but such phosphorylated insulins reduce hyperglycemia when administered to diabetic subjects without inducing hypoglycemia.

Abstract: Amino acid residue sequence and ternary structure of receptor domains for insulin and IGF-I molecules.

Abstract: A novel method for refolding reduced IGF-I is provided in accordance with the present invention. The method is carried out by providing additional positive charge at the amino terminal end of the IGF-I molecule by the presence of a short leader sequence. The additional positive charge on the met-end of the IGF-I molecule has been found to enable recombinant IGF-I to refold simply by stirring solubilized inclusion body protein for between 2-16 hours, or overnight. The present invention also provides a novel fusion protein intermediate of IGF-I attached to the positively charged leader sequence.

Abstract: A process for obtaining a material possessing glucose tolerance factor activity and capable of binding with insulin including the steps of (i) contacting a eukaryotic cell mass with a mixture of water and alcohol to form a water phase and an alcohol phase; (ii) separating the alcohol phase from the water phase and isolating a water phase extract; and (iii) subjecting the water phase extract to gel exclusion chromatography, eluting all material having a molecular weight of approximately 720 to 1120 to obtain a material possessing glucose tolerance factor activity and capable of binding with insulin.

Abstract: Basic proteins are isolated from protein mixtures which contain such basic proteins and which are obtained by enzymatic clevage of proinsulin and/or its derivatives of natural, semisynthetic or genetic engineering origin by ion exchanger chromatography on strongly acid cation exchangers.

Abstract: The invention relates to a process for the isolation and purification of hirudin from complex and salt-containing solutions by hydrophobic chromatography, using as stationary phase porous adsorber resins and as mobile phase organic solvents which are miscible with water.