Combination therapy of growth hormone antagonist and anticancer composition

Abstract

A composition for treating a disease or condition responsive to a human growth hormone receptor antagonist, comprising a modified human growth hormone receptor antagonist; and an anti-cancer composition. A method for treating cancer with a human growth hormone antagonist, comprising pre-screening a patient by analyzing a tumor biopsy to confirm the presence of cancer and the presence of certain predetermined factors indicative of responsiveness to a human growth hormone antagonist; and treating the patient with an effective amount of a composition comprising a modified human growth hormone receptor antagonist and an anti-cancer composition.

Description

[Technical field]

[0001] Sequence Listing Reference A sequence listing in computer readable format (CRF) has been filed. The sequence listing is in an ASCII text (.txt) file entitled SEQIDNOS_1_4_ST25_CIP1.txt, created on August 20, 2021, and is 6KB in size. The sequence listing is incorporated by reference as if fully set forth herein. [Background technology]

[0002] background The disclosed inventive technology relates generally to compositions and methods for treating cancer and other diseases, and more specifically to combination therapies for treating cancer that include a growth hormone antagonist in combination with at least one anti-cancer composition, agent, or drug, which combination therapy prevents the development of drug resistance.

[0003] Human growth hormone (hGH or GH), also known as somatotropin or somatopropin, is a peptide hormone that stimulates growth, cell reproduction, and regeneration in humans and other animals. Growth hormone is a type of mitogen specific to only certain types of cells; it is a single-chain polypeptide of 191 amino acids that is synthesized, stored, and secreted by somatotropic cells in the lateral wings of the anterior pituitary gland. Acromegaly is a syndrome that occurs when the anterior pituitary gland produces too much GH after the closure of the epiphyseal plate during puberty. Excessive production of GH before the closure of the epiphyseal plate results in gigantism (or gigantism). There are many disorders in which pituitary GH production is increased, but the most common involves tumors called pituitary adenomas that originate from a different type of cell (somatotroph cells). Acromegaly most commonly affects middle-aged adults and, if untreated, can cause severe disfigurement, morbidity, and early death. Due to its nature and slow progression, the disease is difficult to diagnose in the early stages and is often overlooked for years until visible changes occur, especially to the face.

[0004] Receptors are protein molecules that are usually embedded in the plasma membrane surface of cells and receive chemical signals from outside the cell. When such chemical signals bind to receptors, they trigger some cellular / tissue response, for example, a change in the electrical activity of the cell. In this sense, receptors are protein molecules that recognize and respond to endogenous chemical signals. Agonists, such as human growth hormone, are chemical compositions that bind to receptors and activate the receptor to cause a biological response. Whereas agonists cause an action, antagonists block the action of agonists, and inverse agonists cause an action opposite to that of agonists. Receptor antagonists are a type of receptor ligand or drug that binds to receptors and blocks or attenuates the response mediated by agonists, rather than causing a biological response itself. These compositions are sometimes called blockers, and examples include alpha blockers, beta blockers, and calcium channel blockers.

[0005] In pharmacology, an antagonist has affinity but no potency for its cognate receptor, and binding disrupts the interaction, inhibiting the function of an agonist or inverse agonist at the receptor. Antagonists mediate their action by binding to the receptor's active site (orthosteric) or other sites (allosteric), or interact at a unique binding site that is not normally involved in the biological regulation of the receptor's activity. Antagonist activity may be reversible or irreversible depending on the lifetime of the antagonist-receptor complex, which depends on the nature of the antagonist-receptor binding. Most drug antagonists exert their potency by competing with endogenous ligands or substrates for structurally defined binding sites on the receptor. By definition, antagonists do not display potency to activate the bound receptor and do not retain the ability to activate the receptor. However, once bound, antagonists inhibit the function of agonists, inverse agonists, and partial agonists.

[0006] Growth hormone receptor antagonists, such as the product pegvisomant (sold under the trademark SOMAVERT®), are used to treat acromegaly. Such compositions are used when the pituitary tumor causing acromegaly cannot be controlled by surgery or radiation and the use of somatostatin analogues has been unsuccessful. Pegvisomant is usually delivered as a powder that is mixed with water and injected subcutaneously.

[0007] PEGylation is the process of attaching polyethylene glycol (PEG) polymer chains, both covalently and non-covalently, to molecules and macrostructures such as drugs, peptides, antibody fragments, or therapeutic proteins. PEGylation is routinely accomplished by incubating reactive derivatives of PEG with the target molecule, resulting in changes in physicochemical properties, including changes in molecular size and molecular charge. These physical and chemical changes can increase the systemic residence of the therapeutic agent, affect the binding affinity of the therapeutic moiety to cellular receptors, and alter absorption and distribution patterns. Covalent attachment of PEG to drugs or therapeutic proteins can also "mask" the drug from the host's immune system (i.e., decrease immunogenicity and antigenicity), increase the hydrodynamic size (i.e., size in solution) of the drug, and increase circulation time by decreasing renal clearance. PEGylation can also impart water solubility to hydrophobic drugs and proteins.

[0008] PEGylation, by increasing the molecular weight of a molecule, can confer several important pharmacological advantages compared to the unmodified form of the molecule, including: (i) improved drug solubility, (ii) reduced dosing frequency without compromising efficacy and potentially reduced toxicity, (iii) increased circulatory life, (iv) improved drug stability, and (v) increased protection from proteolytic degradation. PEGylated drugs also offer commercial advantages, including: (i) opportunities for new delivery forms and dosing regimens; (ii) extended patent life for already approved drugs. PEG is a particularly attractive polymer for conjugation, and the specific properties of the PEG moiety that are relevant for pharmaceutical applications include: (i) water solubility, (ii) high mobility in solution, (iii) lack of toxicity and low immunogenicity, and (iv) altered distribution in the body.

[0009] The addition of high molecular weight polyethylene glycol (PEG) to proteins has previously been shown to increase the in vivo half-life of these proteins by reducing renal excretion in a size-dependent manner. The addition of PEG also reduces protein immunogenicity and decreases aggregation and protease cleavage [1]-[2]. Several known PEGylated proteins have been approved for therapeutic use by the USFDA, including hormones, cytokines, antibody fragments, and enzymes [1], [3]-[4]. Thus, there is a continuing need for further development of PEGylated therapeutics, especially for use in the treatment of diseases responsive to the use of GH receptor antagonists or other receptor antagonists.

[0010] Evidence from human studies [5], animal studies [6], and cancer cell line studies [6] supports a role for growth hormone (GH) in carcinogenesis. The relationship between GH receptor (GHR) expression and function has been examined in the following human cancers: breast, liver, prostate, colon, melanoma, pancreatic, endometrial, meningioma, neuroblastoma, glioma, lung, and gastric cancer [6]. Both in vitro studies using cancer cell lines and in vivo studies using human tumor xenografts in nude mice have shown that attenuating the action of GH can have anticancer effects. The action of GH against cancer has been shown to be inhibited by reducing GH receptor (GHR) expression or by using the GHR antagonist pegvisomant [6]. There is also in vivo and in vitro evidence that attenuating the action of prolactin (PRL) often has anticancer effects against breast and prostate cancers [7].

[0011] The use of GHR antagonists in cancer therapy requires that cancer cells express either the GHR or the prolactin receptor (PRLR), or both. GHR and PRLR expression levels have been measured in 60 cancer cell lines from nine human cancer types: breast, CNS, colon, leukemia, melanoma, non-small cell lung, ovarian, prostate, and kidney cancers.[8] High GHR expression levels were obtained in most cell lines from all cancer types except colon cancer and leukemia. PRLR expression levels were high in most cell lines from all cancer types except leukemia and prostate cancer.

[0012] Chemotherapy is the most common form of cancer treatment, but cancers often acquire resistance to chemotherapy over time [9]. This drug resistance is the main cause of cancer death. The influence of GH on the development of chemotherapy resistance has been recently reviewed

[10] . Studies have examined the effect of GH / GHR signaling on anticancer drug resistance using GH, GHR knockout mice (GHRKO), and the hGHR antagonist pegvisomant

[10] . In vitro studies with cancer cell lines have shown that GH promotes anticancer drug resistance by protecting cells from apoptosis, stimulating epithelial-mesenchymal transition (EMT), stimulating the expression of ATP-binding cassette (ABC) transporters (drug efflux pumps), and promoting cancer stem cells. There is also evidence that PRL promotes cancer drug resistance by upregulating ABC transporters

[11] and promoting cancer stem cell-like properties

[12] .

[0013] Pegvisomant has been shown to reverse GH-stimulated resistance of breast cancer cells to doxorubicin in vitro in cell proliferation assays

[13] . Pegvisomant has also been shown to reverse GH-promoted resistance to doxorubicin-induced apoptosis in cell culture assays

[14] . A novel GHR antagonist (referred to herein as "Compound G") has been prepared that binds to the GHR more tightly than pegvisomant (see U.S. Patent No. 10,874,717). Unlike pegvisomant, Compound G also binds to the PRLR tightly (see U.S. Patent Application No. 16 / 903,644, which is incorporated herein by reference in its entirety). The ability of Compound G to reverse GH-stimulated drug resistance both in vitro and in vivo is described herein. Summary of the Invention

[0014] The following provides a summary of certain exemplary embodiments of the disclosed inventive subject matter. This summary is not an extensive overview and is not intended to identify key or critical aspects or elements of the disclosed inventive subject matter or to delineate the scope thereof. It should be understood, however, that the use of indefinite articles in the language used to describe and claim the disclosed inventive subject matter is not intended to limit the described inventive subject matter in any way. Rather, the use of "a" or "an" should be construed to mean "at least one" or "one or more."

[0015] One embodiment of the disclosed technology provides a composition and method for treating a disease or condition responsive to a human growth hormone receptor antagonist. The composition includes a modified human growth hormone receptor antagonist and an anti-cancer composition. The method for treating a disease or condition responsive to a human growth hormone receptor antagonist includes administering an effective amount of the composition to a patient. The disease or condition responsive to a human growth hormone receptor antagonist can be a cancer expressing a predetermined level of growth hormone receptor (GHR); a predetermined level of prolactin receptor (PRLR); a predetermined level of both GHR and PRLR; a predetermined level of ATP-binding cassette (ABC) transporter; or a predetermined level of epithelial-mesenchymal transition (EMT) mediator. The disease or condition responsive to a human growth hormone receptor antagonist may be breast cancer, central nervous system cancer, melanoma, non-small cell lung cancer, ovarian cancer, prostate cancer, renal cancer, pancreatic cancer, endometrial cancer, meningioma, colorectal cancer, colon cancer, neuroblastoma, gastric cancer, liver cancer, lymphoma, combinations thereof, or other cancers expressing amounts of GHR, PRLR, ABC transporters, EMT mediators, or combinations thereof.

[0016] The modified human growth hormone receptor antagonist may include human growth hormone receptor antagonist G120K in which two amino acids of human growth hormone receptor antagonist G120K have been changed to cysteines, the two amino acids changed to cysteines being T142 and H151; and a polyethylene glycol molecule conjugated to each substituted cysteine ​​of human growth hormone receptor antagonist G120K-T151C-T142C, the polyethylene glycol molecule conjugated to the two amino acids changed to cysteines being two 4.5 kDa branched polyethylene glycols each containing three carboxylate anions. Human growth hormone receptor antagonist G120K may have the DNA sequence of SEQ ID NO:1 and the amino acid sequence of SEQ ID NO:2. Human growth hormone receptor antagonist G120K-H151C-T142C may have the DNA sequence of SEQ ID NO:3 and the amino acid sequence of SEQ ID NO:4. The following amino acid substitutions may be made to prevent binding to the prolactin receptor: H18D, H21N, R167N, K168A, D171S, K172R, E174S, and I179T, and the modified composition may be adapted and used to treat acromegaly. The polyethylene glycol molecule is prepared by stepwise organic chemistry and may be a substantially pure single compound, or the polyethylene glycol molecule may be a branched structure. The polyethylene glycol molecule may include a maleimide group for conjugation with free sulfhydryl groups.

[0017] The anti-cancer composition may be an alkylating agent, an antimetabolite, a plant alkaloid, an antitumor antibiotic, or a combination thereof. The alkylating agent may be chlorambucil, cyclophosphamide, thiotepa, busulfan, cisplatin, or a combination thereof. The antimetabolite may be gemcitabine, 5-fluorouracil, 6-mercaptopurine, cytarabine, or a combination thereof. The plant alkaloid may be vincristine, paclitaxel, etoposide, irinotecan, or a combination thereof. The antitumor antibiotic may be doxorubicin, dactinomycin, mitoxantrone, idarubicin, or a combination thereof. The anti-cancer composition may be a targeted therapy including vemurafenib or a similar agent.

[0018] Another embodiment of the disclosed technology provides another composition and method for treating a disease or condition responsive to a human growth hormone receptor antagonist, the composition includes a modified human growth hormone receptor antagonist, the human growth hormone receptor antagonist includes human growth hormone receptor antagonist G120K, two amino acids of the human growth hormone receptor antagonist G120K are changed to cysteine, the two amino acids changed to cysteine ​​are T142 and H151, and a polyethylene glycol molecule conjugated to each substituted cysteine ​​of the human growth hormone receptor antagonist G120K-H151C-T142C, the polyethylene glycol molecules conjugated to the two amino acids changed to cysteine ​​are two 4.5 kDa branched polyethylene glycols each containing three carboxylate anions, the polyethylene glycol molecules are prepared by stepwise organic chemistry, are substantially pure single compounds, and the polyethylene glycol molecules are branched structures, and an anti-cancer composition. A method for treating a disease or condition responsive to a human growth hormone receptor antagonist comprises administering to a patient an effective amount of the composition.

[0019] The disease or condition responsive to the human growth hormone receptor antagonist may be a cancer that expresses a level of growth hormone receptor (GHR); a level of prolactin receptor (PRLR); a level of both GHR and PRLR); a level of ATP-binding cassette (ABC) transporter; or a level of epithelial-mesenchymal transition (EMT) mediator. The disease or condition responsive to the human growth hormone receptor antagonist may be breast cancer, central nervous system cancer, melanoma, non-small cell lung cancer, ovarian cancer, prostate cancer, kidney cancer, pancreatic cancer, endometrial cancer, meningioma, colorectal cancer, colon cancer, neuroblastoma, gastric cancer, liver cancer, lymphoma, combinations thereof, or any other cancer that expresses a predetermined amount of GHR, PRLR, ABC transporter, EMT mediator, or combinations thereof. The human growth hormone receptor antagonist G120K may have the DNA sequence of SEQ ID NO:1 and the amino acid sequence of SEQ ID NO:2. The human growth hormone receptor antagonist G120K-H151C-T142C may have the DNA sequence of SEQ ID NO:3 and the amino acid sequence of SEQ ID NO:4. The following amino acid substitutions: H18D, H21N, R167N, K168A, D171S, K172R, E174S, and I179T may be substituted to prevent binding to the prolactin receptor, and this modified composition may be adapted and used to treat acromegaly. The polyethylene glycol molecule may contain a maleimide group for coupling with free sulfhydryl groups.

[0020] The anti-cancer composition may be an alkylating agent, an antimetabolite, a plant alkaloid, an antitumor antibiotic, or a combination thereof. The alkylating agent may be chlorambucil, cyclophosphamide, thiotepa, busulfan, cisplatin, or a combination thereof. The antimetabolite may be gemcitabine, 5-fluorouracil, 6-mercaptopurine, cytarabine, or a combination thereof. The plant alkaloid may be vincristine, paclitaxel, etoposide, irinotecan, or a combination thereof. The antitumor antibiotic may be doxorubicin, dactinomycin, mitoxantrone, idarubicin, or a combination thereof. The anti-cancer composition may be a targeted therapy including vemurafenib or a similar agent.

[0021] Yet another embodiment of the disclosed technology provides a method of treating cancer with a human growth hormone antagonist comprising pre-screening a patient by analyzing a tumor biopsy to confirm the presence of cancer and the presence of certain predetermined factors indicative of responsiveness to a human growth hormone antagonist; and treating the patient with an effective amount of a composition comprising a modified human growth hormone receptor antagonist and an anti-cancer composition. The particular predetermined factors may include predetermined levels of GHR, PRLR, ABC transporters, EMT mediators, insulin-like growth factor-1 (IGF-1), IFN-binding protein-3 (IGFBP3), suppressor of cytokine signaling (SOCS_-1, -2, -3, and cytokine-inducible SH2-containing protein (CISH). The cancer may be breast cancer, central nervous system cancer, melanoma, non-small cell lung cancer, ovarian cancer, prostate cancer, kidney cancer, pancreatic cancer, endometrial cancer, meningioma, colorectal cancer, colon cancer, neuroblastoma, gastric cancer, liver cancer, lymphoma, combinations thereof, or any other cancer that expresses a predetermined amount of the predetermined factor.

[0022] The modified human growth hormone receptor antagonist includes human growth hormone receptor antagonist G120K, wherein two amino acids of human growth hormone receptor antagonist G120K are changed to cysteines, the two amino acids changed to cysteines being T142 and H151; and a polyethylene glycol molecule conjugated to each substituted cysteine ​​of human growth hormone receptor antagonist G120K-H151C-T142C, wherein the polyethylene glycol molecules conjugated to the two amino acids changed to cysteines are two 4.5 kDa branched polyethylene glycols each containing three carboxylate anions, the polyethylene glycol molecules being prepared by stepwise organic chemistry and being a substantially pure single compound, and the polyethylene glycol molecules being a branched structure. The human growth hormone receptor antagonist G120K may have the DNA sequence of SEQ ID NO:1 and the amino acid sequence of SEQ ID NO:2, and the human growth hormone receptor antagonist G120K H151C-T142C may have the DNA sequence of SEQ ID NO:3 and the amino acid sequence of SEQ ID NO:4. The polyethylene glycol molecule may be prepared by stepwise organic chemistry and may be a substantially pure single compound, or the polyethylene glycol molecule may be a branched structure. The polyethylene glycol molecule may include a maleimide group for conjugation with free sulfhydryl groups.

[0023] The anti-cancer composition may be an alkylating agent, an antimetabolite, a plant alkaloid, an antitumor antibiotic, or a combination thereof. The alkylating agent may be chlorambucil, cyclophosphamide, thiotepa, busulfan, cisplatin, or a combination thereof. The antimetabolite may be gemcitabine, 5-fluorouracil, 6-mercaptopurine, cytarabine, or a combination thereof. The plant alkaloid may be vincristine, paclitaxel, etoposide, irinotecan, or a combination thereof. The antitumor antibiotic may be doxorubicin, dactinomycin, mitoxantrone, idarubicin, or a combination thereof. The anti-cancer composition may be a targeted therapy including vemurafenib or a similar agent. [Brief description of the drawings]

[0024] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication containing color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0025] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate one or more exemplary embodiments of the disclosed inventive subject matter and, together with the general description given above and the detailed description given below, serve to explain the principles of the disclosed subject matter, wherein: [Figure 1] Figure 1 is a bar graph depicting the expression levels of GHR mRNA in multiple cell lines derived from 37 different cancer cells, listed along the x-axis of the graph; [Diagram 2]Figures 2A-2C are plots for two breast cancer subtypes (triple-negative and HER2-enriched) showing the difference in patient overall survival (in months) for cancers with low and high levels of growth hormone receptor (GHR) expression. Figure 2A contains data for subtype HER2-enriched (ER-PR-HER+) breast cancer (n=96), Figure 2B contains data for subtype triple-negative breast cancer (n=405), and Figure 2C contains data for subtype triple-negative breast cancer / chemotherapy treatment (n=227); [Diagram 3] Figures 3A-3D are graphs showing the relationship between specific gene expression in the Cancer Genome Atlas (TCGA) breast cancer database and overall survival of breast cancer (BC) patients (ungrouped / overall), where data from patients with high GHR expression (Figure 3A) or high PRLR expression (Figure 3B) showed poorer survival compared to low GHR expression or low PRLR expression groups, and breast cancer patients with high expression of both GHR and PRLR (Figure 3C) or high expression of GH, GHR, and PRLR (Figure 3D) had significantly poorer survival compared to the corresponding low expression cohorts of the same gene sets (processed using GEPIA2); [Figure 4] Figure 4A is a heat map showing the Spearman correlation coefficient between GHR expression and ABC-type multidrug transporter expression across 40 different cancer types from human patients in The Cancer Genome Atlas (TCGA) database, which shows a consistent positive (red) correlation between GHR expression and ABC-type multidrug transporters. Figure 4B is a heat map showing the Spearman correlation coefficient between GHR expression and known EMT marker expression across 40 different cancer types from human patients in the TCGA database, which shows a consistent positive (red) correlation between GHR and EMT markers across the majority of cancer types. [Diagram 5]Figures 5A(1)-5A(6) show the effect of GHR antagonists on the expression of six ABC transporters in melanoma cell lines, with Figure 5A(1) containing data for MALME-3M-ABCB1, Figure 5A(2) containing data for MALME-3M-ABCB8, Figure 5A(3) containing data for MALME-3M-ABCC-1, Figure 5A(4) containing data for MALME-3M-ABCC2, Figure 5A(5) containing data for MALME-3M-ABCG1, and Figure 5A(6) containing data for MALME-3M-ABCG2; Figure 5B(1)-Figure 5B(9) show the effect of GHR antagonists on the expression of nine EMT markers in melanoma cell lines, where Figure 5B(1) contains data for MALME-3M-FGFBP1, Figure 5B(2) contains data for MALME-3M-CDH1, Figure 5B(3) contains data for MALME-3M-CLDN1, Figure 5B(4) contains data for MALME-3M-ZEB1, Figure 5B(5) contains data for MALME-3M-CDH2, Figure 5B(6) contains data for MALME-3M-EPAS1, Figure 5B(7) contains data for MALME-3M-SNAI1, Figure 5B(8) contains data for MALME-3M-SNAI2, and Figure 5B(9) contains data for MALME-3M-VIM; [Figure 6] Figures 6A-6D show the effect of GHR antagonists (Peg = pegvisomant, G = compound G) on basement membrane invasion assay (in vitro) (Figure 6A) using three pancreatic cancer cell lines (n = 3, * indicates p < 0.05), GH increases tumor cell invasion rate, but GHR antagonists significantly inhibit tumor cell invasion rate, Figure 6B contains data of invasion assay-BxPC3, Figure 6C contains data of invasion assay-PANC1, Figure 6B contains data of invasion assay-LTPA; [Figure 7]Figures 7A, 7B, and 7C(1)-7C(8) show the effects of GHR antagonists [Peg = pegvisomant, G = compound G] on the drug efflux rate (Figure 7A), drug retention rate (Figure 7B), and fluorescence images after loading a fluorescent drug surrogate (DiOC2) into pancreatic cancer cells (n = 3, * indicates p < 0.05) (Figures 7C(1)-7C(8)). GH increased the drug efflux rate (reduced drug retention in tumor cells), whereas GHR antagonists significantly inhibited it; [Figure 8] Figure 8 shows the viability of pancreatic cancer cells when incubated in the presence of anticancer drugs and either PBS, GH, GH + pegvisomant, or GH + compound G, where the anticancer drugs used were doxorubicin (doxo), erlotinib (erlo), or gemcitabine (gemc), and the control was DMSO; [Figure 9]FIG. 9A is a graph showing the effect of various treatments on the growth (tumor volume) of human pancreatic cancer (PANC1) xenografts implanted in male nude mice, with hGHR antagonists pegvisomant (peg) and compound-G (G) treated at 10 mg / kg / day, while gem20 = gemcitabine was treated at 20 mg / kg every 3 days. All treatments were initiated on day 17 and were by intraperitoneal injection, and gemcitabine, pegvisomant, or compound G, when used alone, were effective in reducing tumor growth rate; however, the combination of gem20+peg or gem20+G was significantly more effective than gem20 or peg or G alone; Figure 9B is a graph showing the effect of various treatments on the growth (tumor volume) of human pancreatic cancer (PANC1) xenografts implanted in male nude mice, with the hGHR antagonist compound-G (G) treated at 10 mg / kg / day, gem20=gemcitabine treated at 20 mg / kg every 3 days, and gem80=gemcitabine treated at 80 mg / kg every 3 days, and all treatments were initiated on day 17 and were by intraperitoneal injection. Both doses of gemcitabine or compound-G were effective in reducing tumor growth rate when used as monotherapy; however, the combination of gem20+G or gem80+G was significantly more effective than gem20 or gem80 or G alone, Gem20+G was more effective than Gem20, Gem80+G was more effective than Gem80 alone, and Gem20+G was more effective than Gem80 alone; and [Figure 10]Figure 10 is a graph showing the effect of various treatments on the growth (tumor volume) of human pancreatic cancer (PANC1) xenografts implanted in female nude mice, where the hGHR antagonist is compound-G (G) treated at 10 mg / kg / day, while gem20 = gemcitabine treated at 20 mg / kg every 3 days, or gem80 = gemcitabine treated at 80 mg / kg every 3 days. All treatments were initiated on day 17 and were by intraperitoneal injection. Both doses of gemcitabine or compound G were effective in reducing tumor growth rate when used as monotherapy; however, the combination of gem20+G or gem80+G was significantly more effective than gem20 or gem80 or G alone, gem20+G was more effective than gem20, but gem80+G was more effective than gem80 alone, and gem20+G was as effective as gem80 alone. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0026] Examples will now be described with reference to the figures. Reference numerals are used throughout the detailed description to refer to various elements and structures. Although the following detailed description contains many specific details for purposes of explanation, those skilled in the art will appreciate that many variations and modifications to the following details are within the scope of the disclosed inventive subject matter. Thus, the following embodiments are described without any loss of generality to, and without imposing limitations on, the claimed subject matter.

[0027] The following abbreviations used throughout this application have the following specific meanings: dPEG®-A is MAL-dPEG® 12 -Tris(-dPEG®) 24-acid)3 (Quanta BioDesign #1145). Compound G or G refers to hGH HR antagonist hGH-G120K in which T142 and H151 have been modified to cysteines and both added cysteines have been conjugated to dPEG®-A. Compound D or D refers to hGH HR antagonist hGH-G120K in which T142 has been modified to cysteine ​​and the added cysteine ​​has been conjugated to a 40 kDa branched polyethylene glycol. Peg refers to pegvisomant and Dox refers to doxorubicin. The disclosed technology includes compositions and methods for treating cancer patients identified by expression of GHR, PRLR, selected ABC drug efflux pumps, selected EMT modulators, IGF-1, IGFBP3, SOCS-1, or CISH. The treatment comprises administering to the patient an effective amount of a chemotherapeutic agent in combination with an effective amount of Compound G.

[0028] GHR expression in cancer cell lines The efficacy of hGHR antagonists in cancer therapy, either by themselves or in combination with anticancer drugs, is closely related to the expression of hGHR by a particular cancer. If the hGHR antagonist is also an antagonist of PRLR, the PRLR expression level will also determine the sensitivity of the cancer to this treatment. Previously, it was observed that most of 60 cell lines from nine cancer types express high levels of either hGHR, PRLR, or both receptors [5]. Analysis of GHR mRNA expression levels from 37 cancer types, including 4 to 131 cell lines for each cancer type (see Figure 1), showed that almost all cancer types have high hGHR expression in at least a subset of the individual cancers tested.

[0029] hGHR expression levels across multiple human patient datasets correlate with decreased patient survival in HER2-enriched and triple-negative breast cancers (see Figures 2A-2C). Figures 3A-3D show that breast cancer patients with high levels of both GHR+PRLR expression or GH+GHR+PRLR expression have poorer survival than breast cancer patients with high levels of GH or GHR or PRLR expression alone.

[0030] GHR expression and expression of ABC transporters and EMT markers Figure 4A shows the correlation between GHR expression and ABC transporter expression across 40 different cancer cell types in all patients in the TCGA database. Figure 4B shows the same correlation between GHR expression and EMT mediator expression. These figures show that chemoresistance is associated with GHR expression, since upregulation of ABC transporters and EMT mediators leads to chemoresistance.

[0031] The effect of GHR antagonists on the expression of six ABC transporters in melanoma cell lines is shown in Figure 5A(1)-5A(6). When used as monotherapy, compound G significantly reduces the expression of four of the six ABC transporters. When GH or doxorubicin is present, compound G also reduces the expression of the four ABC transporters. When both GH and doxorubicin are added, compound G greatly reduces the expression of all six ABC transporters. This observation is due to the fact that the presence of GH or doxorubicin, or both GH and doxorubicin, enhances the expression of ABC transporters.

[0032] The effect of hGHR antagonists on the expression of nine EMT markers in melanoma cell lines is shown in Figure 5B(1)-5B(9). When used as monotherapy, compound G significantly reduces the expression of multiple EMT mediators. In the presence of GH or doxorubicin, or both, compound G also reduces the expression of multiple EMT mediators, presumably because the expression of EMT mediators is significantly enhanced in the presence of GH or doxorubicin, or both GH and doxorubicin.

[0033] Figures 6A-6D show the effect of the hGHR antagonists Compound G and pegvisomant in a basement membrane invasion assay using three pancreatic cancer cell lines. This assay quantifies the ability of cells to migrate across a membrane, a characteristic of cells that have undergone an epithelial to mesenchymal transition. In all three cell lines, the addition of GH increased cell migration. This effect of GH is inhibited by the addition of pegvisomant or Compound G.

[0034] Effect of hGHR antagonists on drug efflux The effect of hGHR antagonists on drug efflux rate from pancreatic cancer cells using DiOC2 dye as a drug surrogate is shown in Figures 7A-7C. Referring to Figure 7A, after 120 minutes, the efflux rate with GH added is about 4 times higher than without (PBS). In the presence of GH + pegvisomant or GH + compound G, the efflux rate is significantly suppressed by about 2 times compared to GH alone. Figure 7B shows the effect of hGHR antagonists on drug retention from pancreatic cancer cells. In the PBS control, about 85% of the drug is retained after 120 minutes. Retention after 120 minutes decreases to ~54% in the presence of GH, but increases to ~83% in the presence of GH + compound G or GH + pegvisomant. Finally, Figures 7C(1)-7C(8) provide fluorescence images of labeled agents after loading into pancreatic cancer cells and incubating for 0 and 120 min in PBS (Figures 7C(1)-7C(2)), GH (Figures 7C(3)-7C(4)), GH + pegvisomant (Figures 7C(5)-7C(6)), or GH + Compound G (Figures 7C(7)-7C(8)). In Figures 7C(1)-7C(8), the agent remaining (fluorescence) after 120 min was compared to its own 0 min control, as the fluorescence of cells at 0 min differed for the four different conditions. As is evident from the images, cells treated with GH + Compound G retained the most drug.

[0035] Figure 8 shows the viability of pancreatic cancer cells when incubated in the presence of anticancer drugs plus buffer (PBS), GH, GH + pegvisomant, or GH + compound G. The anticancer drugs were doxorubicin (doxo), erlotnib, or gemcitabine (gemc), and the control was the anticancer drug vehicle, DMSO. In all cases, the addition of GH increased cell viability. However, the addition of pegvisomant or compound G to GH reduced viability below that of PBS in all cases.

[0036] Effect of GHR antagonist plus anticancer drug on pancreatic tumor xenografts The specific volumes of pancreatic tumor xenografts implanted in male nude mice after treatment with hGHR antagonists (10 mg / kg / day), gemcitabine (20 mg / kg / 3 days), and the combination of antagonists plus gemcitabine are shown in Figure 9A. Treatment began on day 17. Both hGHR antagonists (pegvisomant and compound G) significantly reduced tumor volume compared to the PBS control by day 30 (13 days after treatment initiation). However, tumor volume continued to increase for both of these antagonists. Gemcitabine alone reduced tumor volume more than the hGHR antagonists, but the absolute tumor volume appears to resume an increasing trend by day 43. Gemcitabine + pegvisomant and gemcitabine + compound G combination therapy reduced tumor volume the most. There is no indication that the tumor volume began to increase with the combination therapy after day 43 (day 26 of treatment). The combination of Compound G + gemcitabine reduced tumor volume the most.

[0037] Figure 9B shows the volume of pancreatic tumor xenografts implanted in male nude mice after treatment with 80 mg / kg / 3 days of gemcitabine and compound G (10 mg / kg / day) + 80 mg / kg / 3 days of gemcitabine. Also included in Figure 9B are plots of PBS, compound G (10 mg / kg / day) alone, gemcitabine (20 mg / kg / 3 days) alone, or compound G (10 mg / kg / day) + gemcitabine (20 mg / kg / 3 days) taken from Figure 9A. Gemcitabine alone at 80 mg / kg / 3 days did not significantly reduce the tumor volume obtained with the same drug at 20 mg / kg / 3 days, and with the 80 mg / kg / 3 days dosing regimen, tumor volume appeared to plateau by day 43 (day 26 of treatment). Gemcitabine 80 mg / kg / 3 days plus Compound G 10 mg / kg / 3 days caused the greatest tumor volume reduction under all conditions, and tumor volumes still appeared to be reduced on the last day of the study (day 43 of treatment).

[0038] Figure 10 shows the volume of pancreatic tumor xenografts implanted in female nude mice after treatment with compound G (10 mg / kg / day) alone, gemcitabine (20 or 80 mg / kg / 3 days) alone, compound G (10 mg / kg / day) + 20 mg / kg / 3 days gemcitabine, or compound G (10 mg / kg / day) + 80 mg / kg / 3 days gemcitabine. Treatment began on day 17. After 25 days of administration (42 days after the start of the study), tumor shrinkage was almost the same after administration of 20 mg / kg / 3 days gemcitabine alone or compound G (10 mg / kg / day) alone, but only the gemcitabine-treated mice showed a tendency to increase in tumor volume, indicating the development of chemotherapy resistance. The combination of Compound G (10 mg / kg / day) + Gemcitabine (20 mg / kg / 3 days) dramatically inhibited the increase in tumor volume, which did not appear to trend upward until the end of the study (day 44). Gemcitabine 80 mg / kg / 3 days administered alone produced the same tumor shrinkage effect at day 44 as Compound G (10 mg / kg / day) + Gemcitabine 20 mg / kg / 3 days administered. Compound G (10 mg / kg / day) + Gemcitabine 80 mg / kg / 3 days showed the greatest inhibition of tumor growth, which appeared to decrease even after day 44 (end of study), at which point 3 of 8 mice in the group were tumor-free.

[0039] Personalized Medicine / Precision Medicine Pre-Diagnostic Testing One embodiment of the disclosed technology includes a preliminary molecular analysis of a tumor biopsy sample to determine whether a patient is a suitable candidate for treatment with the disclosed combination therapy. This analysis includes analyzing the expression levels of a predetermined set of genes, and specific changes in the expression levels of these genes correlate with biological activities affected by the disclosed combination therapy. More specifically, identification of elevated expression levels of selected genes is used to identify patients who are suitable candidates for treatment with a GHR antagonist and a cancer therapeutic agent.

[0040] Genes whose expression levels are important indicators of effective responsiveness to the disclosed GHR antagonists and cancer treatments include GHR, PRLR, or both GHR and PRLR. Expression levels in tumor biopsies are measured and quantified by performing diagnostic tests that measure the levels of mRNA encoding these proteins expressed by tumor cells. For example, tumor biopsy samples are processed to isolate mRNA, which is reverse transcribed into cDNA. The amount of cDNA derived from the genes encoding these two receptors can then be measured using a variety of standard assays, including qPCR and gene chip analysis. Patients whose tumors express high levels of GHR, PRLR, or both GHR and PRLR are potential candidates for treatment with the disclosed combination therapy. Alternatively, the levels of these target proteins can be measured using techniques that directly measure the amount of these proteins present in the tumor. This method includes the use of assays such as Western blots and ELISA assays.

[0041] Other genes whose expression levels are important indicators of an effective response to the disclosed hGHR antagonist + cancer therapeutic combination therapy include the major set of ATP-binding cassette (ABC) drug efflux pumps; ABCB1, ABCB5, ABCB8, ABCC1, ABCC2, ABCG1 and ABCG2. As with the target receptors described above, elevated expression levels of at least some of these proteins identify patients who would benefit from the disclosed hGHR antagonist + cancer therapeutic combination therapy. Expression levels of these key genes are determined from samples obtained from patient biopsies using the analytical techniques described above.

[0042] In addition to the drug efflux pump proteins discussed herein, the expression levels of a selected set of genes involved in promoting the deleterious progression of cancer driven by epithelial-mesenchymal transition (EMT) can be measured. The set of key EMT regulators analyzed in the preliminary diagnostic analysis of patient biopsies includes CDH1, CDH2, SNAI1, SNAI2, TGFB1, TGFB2, TGFB3, TGFBR2, TWIST1, TWIST2, VIM, ZEB1 and ZEB2. Elevated expression levels of these genes further identify patients who are candidates for effective treatment with the combination of the disclosed GHR antagonists and cancer therapeutics. The expression levels of these genes will be determined by the above-mentioned analysis methods.

[0043] In addition to the target receptors GHR and PRLR, insulin-like growth factor 1 (IGF-1), insulin-like growth factor binding protein 3 (IGFBP3), suppressor of cytokine signaling (SOCS)-1, -2, -3, and cytokine-inducible SH2-containing protein (CISH) are important genes whose (RNA or protein) expression levels in tumor biopsies (all of the above) or serum (IGF1 and IGFBP3) can be used to identify patients who will respond effectively to treatment with the disclosed GHR antagonists. As with the target receptor proteins, the levels of these GH-inducible downstream signaling factors are determined by gene expression analysis using mRNA gene expression techniques or preferably serum protein quantification techniques. IGF-1, IGFBP3, SOCS-1, -2, -3, and CISH are particularly useful for identifying patients who will be effectively treated by continued administration of the GHR antagonist after the end of combined therapy with the GHR antagonist and a cancer chemotherapeutic agent.

[0044] All literature and similar materials cited in this application, including but not limited to patents, patent applications, articles, books, treatises, web pages, regardless of the form of such literature and similar materials, are expressly incorporated by reference in their entirety. In the event that one or more of the incorporated literature and similar materials differs or conflicts with this application, including but not limited to defined terms, term usage, described techniques, etc., this application controls.

[0045] As mentioned above, and as used herein, the singular forms "a," "an," and "the" refer to both the singular and the plural, unless the context clearly dictates otherwise. As used herein, the term "comprising" is synonymous with "including," "containing," or "characterized by," and is inclusive or open-ended and does not exclude additional, implied elements or method steps. Although many methods and materials similar or equivalent to those described herein can be used, certain preferred methods and materials are described herein. Unless the context dictates otherwise, the recitation of numerical ranges by endpoints includes all numbers subsumed within that range. Furthermore, references to "one embodiment" are not intended to be interpreted as excluding the existence of additional embodiments that also incorporate the recited features. Furthermore, unless expressly stated to the contrary, an embodiment "comprising" or "having" an element or elements having a particular characteristic can include additional elements whether or not they have that characteristic.

[0046] As used throughout this specification, the terms "substantially" and "about" are used to account for and take into account small variations due to processing variations, etc. For example, these terms can refer to ±5% or less, such as ±2% or less, such as ±1% or less, such as ±0.5% or less, such as ±0.2% or less, such as ±0.1% or less, such as ±0.05% or less, and / or 0%.

[0047] Underlined and / or italic headings and subheadings are used for convenience only, are not intended to limit the disclosed subject matter, and are not to be referenced in connection with interpreting the description of the disclosed subject matter. All structural and functional equivalents to the elements of the various embodiments described throughout this disclosure that are known or that later become known to those skilled in the art are expressly incorporated herein by reference and are intended to be encompassed by the disclosed subject matter. Furthermore, nothing disclosed herein is intended to be generally exclusive, regardless of whether such disclosure is expressly set forth in the description above.

[0048] There may be many alternative ways of implementing the disclosed inventive subject matter. The various functions and elements described herein may be divided differently than shown without departing from the scope of the disclosed inventive subject matter. The general principles defined herein may be applied to other implementations. Different numbers of a given module or unit may be employed, different types or varieties of a given module or unit may be employed, given modules or units may be added, and given modules or units may be omitted.

[0049] In the context of this disclosure, the term "plurality" refers to two or more than two. Unless otherwise clearly defined, the orientation or positional relationship indicated by terms such as "upper" and "lower" is based on the orientation or positional relationship as shown in the figures only to facilitate and simplify the description of the present invention, and does not indicate or imply that the referenced device or element must be in a particular orientation or configured or operated in a particular orientation, and therefore should not be construed as limiting the present invention. Terms such as "connected," "mounted," and "fixed" should be understood in a broad sense. For example, "connected" may be a fixed connection, a detachable connection, or an integral connection, a direct connection, or an indirect connection through an intermediate medium. For those skilled in the art, the specific meaning of the above terms in the present invention can be understood according to the specific situation.

[0050] It should be understood that all combinations of the foregoing concepts and additional concepts discussed in more detail herein (provided such concepts are not mutually inconsistent) are contemplated as part of the disclosed inventive subject matter. In particular, all combinations of claimed subject matter appearing at the end of this disclosure are contemplated as part of the disclosed inventive subject matter. The disclosed inventive subject matter has been illustrated by a description of exemplary embodiments, and although the exemplary embodiments have been described in certain details, there is no intention to limit the scope of the appended claims to such details or in any way to such details. Additional advantages and modifications will readily occur to those skilled in the art. Thus, the disclosed inventive subject matter in its broader aspects is not limited to any of the specific details, representative apparatus and methods, and / or illustrative examples shown and described. Thus, departures may be made from such details without departing from the spirit or scope of the general inventive concept.

[0051] The following references form part of this application, and each reference is incorporated herein by reference in its entirety for all purposes. 1. Pasut,G. and Veronese,M.(2012) State of the Art in Pegylation:The Great Versatility Achieved After Forty Years of Research.J.Controlled Release 161,461-472. 2. Parveen,S.and Sahoo,S.K.Nanomedicine:Clinical Applications of Polyethylene Glycol Conjugated to Proteins and Drugs Clin.Pharmacokinet.45,965-988. 3. Alconcel, S.N.S., Baas, A.S. and Maynard, H.D. (2011) FDA-Approved Poly(ethylene glycol)-Protein Conjugate Drugs. Polymer Chemistry 2, 1442-1448. 4. Kling, J. (2013) Pegylation of Biologics: A Multipurpose Solution. Bioprocess International 11, 35-43. 5. Perry, J.K., Wu, Z.-S., Mertani, H.C., Zhu, T., and Lobie, P.E. (2017) "Tumour- Derived Human Growth Hormone as a Therapeutic Target in Oncology" Trends in Endocrinology and Metabolism 28: 587-596. 6. Basu, R., Qian, Y., and Kopchick, J.J. (2018) "Lessons from growth hormone receptor gene-disrupted mice: are there benefits of endocrine defects?" European Journal of Endocrinology 178: R155-R181. 7. Goffin, V. (2017) "Prolactin Receptor Targeting in Breast and Prostate Cancers: New Insights into an Old Challenge" Pharmacology and Therapeutics 179: 111-126. 8. Sustarsic, E.G., Junnila, R.K., and Kopchick, J.J. (2013) "Human Metastatic Melanoma Cell Lines Express High Levels of Growth Hormone Receptor and Respond to GH Treatment" Biochem Biophys Res Commun. 441: 144-150. 9. Bukowski, K., Kciuk, M., and Kontek, R. (2020) "Mechanisms of Multidrug Resistance in Cancer Chemotherapy" Int. J. Mol. Sci. 21, 3233 10. Basu, R., and Kopchick, J.J. (2019) "The Effects of Growth Hormone on Therapy Resistance in Cancer" Cancer Drug Resistance 2: 827-846, 11. Wu, A.M.L., Dalvi, P., Lu, X., Yang, M., Riddick, D.S., et al. (2013) "Induction of multidrug resistance transporter ABCG2 by prolactin in human breast cancer cells" Molecular Pharmacology 83:377-88. 12. Neradugomma, N.K., Subramaniam, D., Tawfik, O.W., Goffin, V., Kumar, T.R., et al. (2014) "Prolactin signaling enhances colon cancer stemness by modulating Notch signaling in a Jak2-STAT3 / ERK manner" Carcinogenesis 35:795-806 13. Zatelli, M.C., Minoia, M., Mole, D., Cason, V., Tagliati, F., Margutti, A., Bondanelli, M., Ambrosio, M.R., and Uberti, E.d (2009) "Growth Hormone Excess Promotes Breast Cancer Chemoresistance" Journal of Clinical Endocrinology and Metabolism 94: 3931-3938. 14. Minoia, M., Gentilin, E., Mole, D., Rossi, M., Filieri, C., Tagliati, F., Baroni, A., Ambrosio, M.R., and Uberti, E.d, Zatelli, M.C. "Growth Hormone Receptor Blockade Inhibits Growth Hormone-Induced Chemoresistance by Restoring Cytotoxic-Induced Apoptosis in Breast Cancer Cells Independently of Estrogen Receptor Expression" Journal of Clinical Endocrinology and Metabolism 97: E907-E916.

Claims

1. 1. A composition for treating a disease or condition responsive to a human growth hormone receptor antagonist, comprising: (a) a modified human growth hormone receptor antagonist; and (b) an anti-cancer composition; A composition comprising:

2. 2. The composition of claim 1, wherein the disease or condition responsive to a human growth hormone receptor antagonist is a cancer expressing a predetermined level of growth hormone receptor (GHR), a predetermined level of prolactin receptor (PRLR), a predetermined level of both GHR and PRLR, a predetermined level of ATP-binding cassette (ABC) transporter, or a predetermined level of an epithelial-mesenchymal transition (EMT) mediator.

3. 2. The composition of claim 1, wherein the disease or condition responsive to a human growth hormone receptor antagonist is cancer, and the cancer is breast cancer, central nervous system cancer, melanoma, non-small cell lung cancer, ovarian cancer, prostate cancer, renal cancer, pancreatic cancer, endometrial cancer, meningioma, colorectal cancer, colon cancer, neuroblastoma, gastric cancer, liver cancer, lymphoma, combinations thereof, or any other cancer that expresses a predetermined amount of GHR, PRLR, ABC transporter, EMT mediator, or combinations thereof.

4. 2. The composition of claim 1, wherein the modified human growth hormone receptor antagonist is (a) human growth hormone receptor antagonist G120K, wherein two amino acids of human growth hormone receptor antagonist G120K are changed to cysteine, the two amino acids changed to cysteine ​​being T142 and H151; (b) a polyethylene glycol molecule conjugated to each substituted cysteine ​​of human growth hormone receptor antagonist G120K-H151C-T142C, wherein the polyethylene glycol molecules conjugated to the two amino acids changed to cysteines are two 4.5 kDa branched polyethylene glycols each containing three carboxylate anions; A composition comprising:

5. 5. The composition of claim 4, wherein the human growth hormone receptor antagonist G120K has the DNA sequence of SEQ ID NO:1 and the amino acid sequence of SEQ ID NO:

2.

6. The composition of claim 4, wherein the human growth hormone receptor antagonist G120K-H151C-T142C has the DNA sequence of SEQ ID NO:3 and the amino acid sequence of SEQ ID NO:

4.

7. 5. The composition of claim 4, wherein the following amino acid substitutions are made: H18D, H21N, R167N, K168A, D171S, K172R, E174S, and I179T, and these mutations function to prevent binding to the prolactin receptor.

8. 8. The composition of claim 7, wherein the composition is adapted for the treatment of acromegaly.

9. 5. The composition of claim 4, wherein the polyethylene glycol molecule is prepared by stepwise organic chemistry, is a substantially pure single compound, and is a branched structure.

10. The composition of claim 4, wherein the polyethylene glycol molecule comprises a maleimide group for conjugation with free sulfhydryl groups.

11. 10. The composition of claim 1, wherein the anti-cancer compound is an alkylating agent, an antimetabolite, a plant alkaloid, an antitumor antibiotic, or a combination thereof.

12. 12. The composition of claim 11, wherein the alkylating agent is chlorambucil, cyclophosphamide, thiotepa, busulfan, cisplatin, or a combination thereof.

13. The composition of claim 11, wherein the antimetabolite is gemcitabine, 5-fluorouracil, 6-mercaptopurine, cytarabine, or a combination thereof.

14. 12. The composition of claim 11, wherein the plant alkaloid is vincristine, paclitaxel, etoposide, irinotecan, or a combination thereof.

15. 12. The composition of claim 11, wherein the antitumor antibiotic is doxorubicin, dactinomycin, mitoxantrone, idarubicin, or a combination thereof.

16. The composition of claim 1 , wherein the anti-cancer composition is a targeted therapy.

17. 17. The composition of claim 16, wherein the targeted therapy comprises vemurafenib.

18. Use of a modified human growth hormone receptor antagonist for the manufacture of a composition according to claim 1.

19. 1. A composition for treating a disease or condition responsive to a human growth hormone receptor antagonist, comprising: (a) a modified human growth hormone receptor antagonist, said modified human growth hormone receptor antagonist comprising: (i) human growth hormone receptor antagonist G120K, wherein two amino acids of human growth hormone receptor antagonist G120K are changed to cysteine, the two amino acids changed to cysteine ​​being T142 and H151; (ii) a polyethylene glycol molecule conjugated to each substituted cysteine ​​of human growth hormone receptor antagonist G120K-H151C-T142C, wherein the polyethylene glycol molecules conjugated to the two amino acids changed to cysteines are two 4.5 kDa branched polyethylene glycols each containing three carboxylate anions, the polyethylene glycol molecules being prepared by stepwise organic chemistry and being substantially pure single compounds, and the polyethylene glycol molecules being branched structures. and a modified human growth hormone receptor antagonist comprising: (b) an anti-cancer composition; A composition comprising:

20. 20. The composition of claim 19, wherein the disease or condition responsive to a human growth hormone receptor antagonist is a cancer expressing a predetermined level of growth hormone receptor (GHR), a predetermined level of prolactin receptor (PRLR), a predetermined level of both GHR and PRLR, a predetermined level of ATP-binding cassette (ABC) transporter, or a predetermined level of an epithelial-mesenchymal transition (EMT) mediator.

21. 20. The composition of claim 19, wherein the disease or condition responsive to a human growth hormone receptor antagonist is cancer, and the cancer is breast cancer, central nervous system cancer, melanoma, non-small cell lung cancer, ovarian cancer, prostate cancer, renal cancer, pancreatic cancer, endometrial cancer, meningioma, colorectal cancer, colon cancer, neuroblastoma, gastric cancer, liver cancer, lymphoma, combinations thereof, or any other cancer that expresses a predetermined amount of GHR, PRLR, ABC transporter, EMT mediator, or combinations thereof.

22. 20. The composition of claim 19, wherein the human growth hormone receptor antagonist G120K has the DNA sequence of SEQ ID NO: 1 and the amino acid sequence of SEQ ID NO:

2.

23. 20. The composition of claim 19, wherein the human growth hormone receptor antagonist G120K-H151C-T142C has the DNA sequence of SEQ ID NO:3 and the amino acid sequence of SEQ ID NO:

4.

24. 20. The composition of claim 19, wherein the following amino acid substitutions are made: H18D, H21N, R167N, K168A, D171S, K172R, E174S, and I179T, and these mutations function to prevent binding to the prolactin receptor.

25. 25. The composition of claim 24, wherein the composition is adapted for the treatment of acromegaly.

26. 20. The composition of claim 19, wherein the polyethylene glycol molecule comprises a maleimide group for conjugation with free sulfhydryl groups.

27. 20. The composition of claim 19, wherein the anti-cancer compound is an alkylating agent, an antimetabolite, a plant alkaloid, an antitumor antibiotic, or a combination thereof.

28. 28. The composition of claim 27, wherein the alkylating agent is chlorambucil, cyclophosphamide, thiotepa, busulfan, cisplatin, or a combination thereof.

29. 28. The composition of claim 27, wherein the antimetabolite is gemcitabine, 5-fluorouracil, 6-mercaptopurine, cytarabine, or a combination thereof.

30. 28. The composition of claim 27, wherein the plant alkaloid is vincristine, paclitaxel, etoposide, irinotecan, or a combination thereof.

31. 28. The composition of claim 27, wherein the antitumor antibiotic is doxorubicin, dactinomycin, mitoxantrone, idarubicin, or a combination thereof.

32. 20. The composition of claim 19, wherein the anti-cancer composition is a targeted therapy.

33. 33. The composition of claim 32, wherein the targeted therapy comprises vemurafenib.

34. Use of a modified human growth hormone receptor antagonist for the manufacture of a composition according to claim 19.

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