Provided in the present invention are a method for constructing a strain for producing a recombinant protein containing unnatural amino acids, and the strain obtained therefrom. The method comprises modifying the expression of a prfA gene contained in the strain to be controllable. The strain constructed by the method of the present invention can efficiently produce in intact protein containing unnatural amino acids, greatly reduce the production of a truncated protein and can also maintain high-speed growth.
An unnatural amino acid may have a structure as shown in formula (1) or may be an enantiomer thereof:
An unnatural amino acid may have a structure as shown in formula (1) or may be an enantiomer thereof:
An unnatural amino acid may have a structure as shown in formula (1) or may be an enantiomer thereof:
Such unnatural amino acid may be used, e.g., in recombinant proteins. A recombinant protein may contain such unnatural amino acid and a protein conjugate prepared from the recombinant protein. The unnatural amino acid can be simple and convenient to prepare, safe in nature, difficult to inactivate when inserted into a protein, and may have a high binding rate with a coupling moiety, and the obtained conjugate is higher in stability. The unnatural amino acid is applicable to numerous fields, especially in preparation of a recombinant protein or a recombinant protein conjugate.
A61K 47/60 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
C07C 235/20 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated having at least one of the singly-bound oxygen atoms further bound to a carbon atom of a six-membered aromatic ring, e.g. phenoxyacetamides having the nitrogen atoms of the carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
C07C 235/78 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of an unsaturated carbon skeleton the carbon skeleton containing rings
C07C 237/12 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups
C07C 271/22 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
C07C 271/54 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to carbon atoms of six-membered aromatic rings with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
C07K 1/00 - General processes for the preparation of peptides
3.
HETEROARYL-CONTAINING UNNATURAL AMINO ACID AND USE THEREOF
Provided is a heteroaryl-containing unnatural amino acid, which is a compound having a structure as represented by formula (I), or an enantiomer thereof. The protein expression level is higher when the heteroaryl-containing unnatural amino acid is used for preparing a recombinant protein, and the synthesis efficiency is also higher when the heteroaryl-containing unnatural amino acid is used for forming a conjugate. Therefore, the heteroaryl-containing unnatural amino acid has good applicability, and has a simple and convenient preparation process, does not require high costs, and is thus suitable for large-scale production. The provided recombinant human interleukin 2-polyethylene glycol conjugate realizes the accurate site-directed coupling of PEG with IL-2 by means of using the unnatural amino acid, thereby prolonging the half-life of IL-2 in vivo and facilitating the immunotherapy of tumors.
A61K 47/60 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
The present invention provides a freeze-dried composition containing anti-HER2 drug conjugate, which comprises ARX788 and one or more of pharmaceutically acceptable lyoprotectant, buffer salts and surfactants, wherein the pH value of the freeze-dried composition is from 5.7 to 6.3. The present invention also provides a freeze-dried preparation containing anti-HER2 drug conjugate and the preparation method thereof. The present invention also provides use of the freeze-dried composition and the freeze-dried preparation. The freeze-dried composition and the freeze-dried preparation provided by the present invention have a very excellent stability, as well as good appearance, short reconstitution time and low moisture content. The product quality is safe and reliable, and the production efficiency is high. The preparation methods of the freeze-dried composition and the freeze-dried preparation provided by the present invention are simple and have low-cost, and they can be used for treating various cancers, and have broad application prospects.
A61K 9/19 - Particulate form, e.g. powders lyophilised
A61K 47/18 - AminesAmidesUreasQuaternary ammonium compoundsAmino acidsOligopeptides having up to five amino acids
A61K 47/26 - Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharidesDerivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
A61K 47/68 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
5.
HUMAN INTERLEUKIN 2-POLYETHYLENE GLYCOL CONJUGATE AND USE THEREOF
A human interleukin 2-polyethylene glycol conjugate and a use thereof. The human interleukin 2-polyethylene glycol conjugate contains recombinant human interleukin 2 containing at least one unnatural amino acid and PEG coupled to the at least one unnatural amino acid. The unnatural amino acid is a compound containing a carbonyl end group having a structure as shown in formula (I) or an enantiomer thereof; PEG is coupled to the at least one unnatural amino acid by forming an oxime bond between the carbonyl end group and PEG containing a hydroxylamine end group. The human interleukin 2-polyethylene glycol conjugate can be used individually or in combination with other antitumor drugs for the treatment of diseases such as solid tumors and hematological tumors.
A61K 47/60 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
The present invention provides a human interleukin 2-polyethylene glycol (PEG) conjugate and an application thereof. The human interleukin 2-PEG conjugate provided in the present invention comprises recombinant human interleukin 2 containing at least one unnatural amino acid and PEG coupled to the at least one unnatural amino acid. The unnatural amino acid is a compound containing a carbonyl end group and having a structure as represented by formula (I) or an enantiomer of the compound. The PEG is coupled to the at least one unnatural amino acid by forming an oxime bond between the carbonyl end group and the PEG containing a hydroxylamine end group. The human interleukin 2-PEG conjugate provided in the present invention can be used alone or in combination with other antitumor drugs for treating diseases such as solid tumors and blood tumors.
A61K 47/60 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
Provided in the present invention are a method for constructing a strain for producing a recombinant protein containing unnatural amino acids, and the strain obtained therefrom. The method comprises modifying the expression of a prfA gene contained in the strain to be controllable. The strain constructed by means of the method of the present invention can efficiently produce a complete protein containing unnatural amino acids, and also can maintain high-speed growth while greatly reducing the production of a truncated protein.
The invention provides an unnatural amino acid, which is a compound with a structure as shown in formula (I) or an enantiomer thereof. The invention further provides use of the unnatural amino acid. Furthermore, the invention further provides a recombinant protein containing the unnatural amino acid and a protein conjugate prepared from the recombinant protein. The unnatural amino acid provided by the invention is simple and convenient to prepare, safe in nature, not easy to be inactivated when being inserted into a protein, and has a high binding rate with a coupling moiety, and the obtained conjugate is higher in stability. The unnatural amino acid provided by the invention is applicable to numerous fields, especially in preparation of a recombinant protein or a recombinant protein conjugate.
A61K 47/54 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
A61K 47/60 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
A61P 5/06 - Drugs for disorders of the endocrine system of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
C07C 235/20 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated having at least one of the singly-bound oxygen atoms further bound to a carbon atom of a six-membered aromatic ring, e.g. phenoxyacetamides having the nitrogen atoms of the carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
C07C 235/78 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of an unsaturated carbon skeleton the carbon skeleton containing rings
C07C 237/12 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups
C07C 271/22 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
C07K 14/61 - Growth hormone [GH], i.e. somatotropin
9.
UNNATURAL AMINO ACID, AND USE THEREOF, RECOMBINANT PROTEIN CONTAINING SAME, AND RECOMBINANT PROTEIN CONJUGATE
Provided is an unnatural amino acid, which is a compound having a structure as shown in formula (I) or an enantiomer thereof. Also provided is the use of the unnatural amino acid. Further provided is a recombinant protein containing the unnatural amino acid and a protein conjugate prepared from the recombinant protein. The unnatural amino acid is easy to prepare and has a good safety; when the unnatural amino acid is inserted into a protein, the protein is not easily inactivated; the unnatural amino acid has a high binding rate to a conjugated moiety, and the stability of the obtained conjugate is higher; and the unnatural amino acid can be used in numerous fields, especially in the preparation of recombinant proteins or recombinant protein conjugates.
C07C 271/22 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
C07C 235/78 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of an unsaturated carbon skeleton the carbon skeleton containing rings
C07C 237/12 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups
C07C 235/20 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated having at least one of the singly-bound oxygen atoms further bound to a carbon atom of a six-membered aromatic ring, e.g. phenoxyacetamides having the nitrogen atoms of the carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
C07K 14/61 - Growth hormone [GH], i.e. somatotropin
A61K 47/54 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
A61K 47/60 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
A61K 38/27 - Growth hormone [GH], i.e. somatotropin
A61P 5/06 - Drugs for disorders of the endocrine system of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
10.
UNNATURAL AMINO ACID AND APPLICATION THEREOF, RECOMBINANT PROTEIN CONTAINING SAME, AND RECOMBINANT PROTEIN CONJUGATE
Provided by the present invention is an unnatural amino acid, which is a compound having a structure as shown in formula (I) or an enantiomer thereof. Further provided by the present invention is an application of the unnatural amino acid. Furthermore, also provided by the present invention is a recombinant protein containing the unnatural amino acid and a protein conjugate prepared from the recombinant protein. The unnatural amino acid provided by the present invention is easy to prepare and has good safety; moreover, said amino acid does not easily lose its activity when inserted into a protein, and the binding rate to a coupling part is high. The obtained conjugate has good stability. The unnatural amino acid provided by the present invention can be applied in many fields, especially in the preparation of a recombinant protein or a recombinant protein conjugate.
C07C 269/00 - Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups the nitrogen atom not being part of nitro or nitroso groups
C07C 269/06 - Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups the nitrogen atom not being part of nitro or nitroso groups by reactions not involving the formation of carbamate groups
C07C 271/22 - Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
C07C 231/12 - Preparation of carboxylic acid amides by reactions not involving the formation of carboxamide groups
C07C 235/74 - Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of a saturated carbon skeleton
C07D 233/56 - Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
C07D 207/46 - Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with hetero atoms directly attached to the ring nitrogen atom
C07C 51/41 - Preparation of salts of carboxylic acids by conversion of the acids or their salts into salts with the same carboxylic acid part
C07C 201/12 - Preparation of nitro compounds by reactions not involving the formation of nitro groups
C07C 205/43 - Compounds containing nitro groups bound to a carbon skeleton the carbon skeleton being further substituted by esterified hydroxy groups having nitro groups or esterified hydroxy groups bound to carbon atoms of six-membered aromatic rings of the carbon skeleton to carbon atoms of the same non-condensed six-membered aromatic ring or to carbon atoms of six-membered aromatic rings being part of the same condensed ring system
C07C 41/16 - Preparation of ethers by reaction of esters of mineral or organic acids with hydroxy or O-metal groups
C07C 43/178 - Unsaturated ethers containing hydroxy or O-metal groups
C07K 14/61 - Growth hormone [GH], i.e. somatotropin
A61K 38/27 - Growth hormone [GH], i.e. somatotropin
A61K 47/60 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
A61K 47/68 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
A61P 5/06 - Drugs for disorders of the endocrine system of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
Provided in the present invention is a freeze-dried composition containing an anti-HER2 drug conjugate, which comprises ARX788 and one or more of a pharmaceutically acceptable freeze-dried protective agent, a buffer salt and a surfactant, wherein the pH value of the freeze-dried composition is 5.7-6.3. Further provided in the present invention are a freeze-dried preparation containing the anti-HER2 drug conjugate and a preparation method therefor. Also provided in the present invention is the use of the freeze-dried composition and the freeze-dried preparation. The freeze-dried composition and freeze-dried preparation provided in the present invention have a good stability, good appearance, short redissolving time, low water content, safe and reliable product quality and high production efficiency. The freeze-dried composition and freeze-dried preparation provided in the present invention have a simple and convenient preparation method and low cost, can be used for treating various cancers, and have broad application prospects.
A61K 9/19 - Particulate form, e.g. powders lyophilised
A61K 47/68 - Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additivesTargeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
A61K 47/18 - AminesAmidesUreasQuaternary ammonium compoundsAmino acidsOligopeptides having up to five amino acids
A61K 47/26 - Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharidesDerivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
patents
