Treatment of head and neck cancer with pharmaceutical compositions comprising nanoparticles containing taxanes

Abstract

The present disclosure relates to methods for treating cancer patients. The methods include administering to the patient a pharmaceutical composition comprising taxane nanoparticles modified with symmetrically branched homopolymers and asymmetrically branched homopolymers. In particular, the present disclosure relates to methods and single-dose unit pharmaceutical packages for treating head and neck cancer (HNC), head and neck squamous cell carcinoma (HNSCC), and other cancers with the pharmaceutical compositions.

Description

[0001] Cross-references to related applications This application claims priority to U.S. Patent Application No. 63 / 548,806, filed February 1, 2024, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0002] This disclosure relates to a pharmaceutical composition comprising nanoparticles and a method of treating head and neck cancer using the pharmaceutical composition, wherein the nanoparticles contain taxanes modified with a surface-modified branched polymer (MBP) or a linear polymer.

[0003] background According to estimates by the Union for International Cancer Control (UICC), approximately 550,000 people are diagnosed with head and neck cancer (HNC) each year. HNC is the sixth most common cancer and accounts for about 5% of all cancers worldwide. About 90% of all HNCs are squamous cell carcinomas of the head and neck (SCCHN), also known as head and neck squamous cell carcinoma (HNSCC). The mortality rate for HNC is approximately 50%. HNC can include cancers of the throat that can occur anywhere in the throat or head and neck region, such as hypopharyngeal and laryngeal cancers, nasopharyngeal cancers, oral and pharyngeal cancers, sinus and nasal cavity cancers, salivary gland cancers, and thyroid cancers. Despite advances in understanding its epidemiology and pathogenesis, survival rates for many types of HNSCC have barely improved in the past 30–40 years.

[0004] Treatment for advanced HNSCC can include a variety of therapies, such as surgery, postoperative chemoradiotherapy (CRT), radiation therapy, chemotherapy, or combinations thereof. The EGFR monoclonal antibody cetuximab is often combined with radiation therapy for HPV-negative HNSCC, where comorbidity prevents the use of cytotoxic chemotherapy, but the proportion of patients with HPV+ oropharyngeal carcinoma continues to increase. For those with programmed cell death ligand 1 (PD-L1) expression on tumor or immune cells, first-line therapy should be combined with the PD-1-directed antibody pembrolizumab, with or without chemotherapy. For those with HPV+ tumors, the effect on survival is greater when chemotherapy is combined with pembrolizumab. In some studies, paclitaxel has also been used as monotherapy or in combination with cisplatin, 5-fluorouracil, and cetuximab for patients with recurrent or metastatic HNSCC. However, many patients are ineligible for or unsuitable for treatment.

[0005] Taxanes, including paclitaxel (in the form of taxol) ® Sold by Bristol-Myers Squibb) and Dorcetsay (in taxotere) ®Paclitaxel (sold by Aventis Pharma SA, FR) is used to treat various cancers, including breast cancer, ovarian cancer, lung cancer, colon cancer, and head and neck cancer. However, its poor water solubility hinders its widespread use. Currently, taxol... ® And its generic drugs use ethanol: cremaphor ® A 1:1 solution of (BASF's polyethoxylated castor oil) was prepared to dissolve the drug. However, cremaphor ® The presence of this substance has been associated with severe hypersensitivity reactions, and therefore requires pharmacological treatment with corticosteroids (e.g., dexamethasone) and antihistamines. Alternatively, conjugated paclitaxel, such as abraxane produced by mixing paclitaxel with human serum albumin, can be used. ® (Abraxis Bioscience, NJ, USA) has eliminated the need for injections of corticosteroids and antihistamines. However, abraxane... ® Undesirable side effects, such as serious cardiovascular events, can occur, including chest pain, cardiac arrest, supraventricular tachycardia, edema, thrombosis, pulmonary embolism, pulmonary embolism, and hypertension. This prevents patients with high cardiovascular risk from using the drug.

[0006] Branched polymers have been used for drug delivery, but those attempts have primarily focused on the chemical attachment of drugs to polymers or the physical encapsulation of such drugs within monomolecules. For example, dendrigrafts and dendritic grafts are thought to physically encapsulate bioactive molecules using monomolecular encapsulation methods, or to encapsulate various drugs using SBP monomolecules to form “dendritic cassettes” (Tomalia et al., Angew. Chem. Int. Ed. Engl., 1990, 29, 138; “Dendrimers and Other Dendritic Polymers”, edited by Frechet and Tomalia, John Wiley & Sons, Ltd., 2001, pp. 387–424). Block copolymers, such as heteroarm polymers (i.e., Y-shaped / AB2-type star polymers) and linear (A)–dendritic (B) block copolymers, have been observed to form stereocomplexes with paclitaxel. However, such branched block copolymers are difficult to prepare and therefore unsuitable for large-scale production.

[0007] Therefore, it is still necessary to modify branched or linear homopolymers with hydrophobic groups. These branched or linear homopolymers spontaneously form stable aggregates when exposed to poorly soluble or water-insoluble drugs. These stable aggregates are suitable for controlled drug delivery and can be used in more effective methods for treating HNSCC.

[0008] Overview This disclosure relates to a method for treating cancer in a subject with corresponding need, the method comprising administering to the subject an effective dose of a first pharmaceutical composition, wherein the first pharmaceutical composition comprises an aggregate comprising: a) a polyoxazoline comprising at least one first end group modified by a hydrophobic portion, wherein the polyoxazoline further comprises a linear portion, a branched portion, or both, the branched portion comprising a symmetrically branched polymer, an asymmetricly branched polymer, or a combination thereof; and the polyoxazoline comprises a monomer to initiator molar ratio in the range of 50:1 to 80:1, and b) taxane, wherein the subject has recurrent or metastatic cancer and has been treated for at least 7 days prior to the first administration of the first pharmaceutical composition with radiotherapy, surgery, chemotherapy, hormone therapy, biotherapy, immunotherapy, checkpoint inhibitor therapy, programmed death-ligand 1 (PD-L1)-based immune checkpoint inhibitor therapy, or a combination thereof. In some embodiments of the method, the aggregate comprises a polyoxazoline to taxane weight ratio of 6:1 to 8:1; and has a size of 70 nm to 90 nm. In some embodiments of the method, the polyoxazoline comprises a second end group containing a functional group modified with ethylenediamine (EDA) or an EDA derivative, wherein the ratio of the second end group to EDA is 1:10. In some embodiments of the method, the EDA derivative is configured to include diethylenetriamine, triethylenetetramine, tetraethylenepentamine, pentaethylenehexamine, polyvinylamine, or tetramethylethylenediamine. In some embodiments of this method, the subject has been treated with at least one drug, including afatinib, albumin-bound paclitaxel or nab-paclitaxel, bleomycin sulfate, carboplatin, cisplatin, cetuximab, docetaxel, hydroxyurea, pembrolizumab, methotrexate, nivolumab, cisplatin (Platinol), paclitaxel, docetaxel, Trexall (methotrexate), a combination of carboplatin and taxol, a combination of TPF (docetaxel, cisplatin, and fluorouracil (FU)), or any of the foregoing combinations. In some embodiments of this method, the subject has been treated with: pembrolizumab as a single agent with a combination positive score (CPS) ≥1; a combination of pembrolizumab, platinum, and fluorouracil; cetuximab; docetaxel; nivolumab; afatinib; or any of the foregoing combinations. In some embodiments of this method, the subject has been treated with: first-line therapy comprising pembrolizumab or a combination of pembrolizumab, platinum, and fluorouracil; and second-line therapy comprising cetuximab, docetaxel, nivolumab, afatinib, or a combination thereof. In some embodiments of this method, the cancer is breast cancer, triple-negative breast cancer, ovarian cancer, lung cancer, NSCLC (non-small cell lung cancer), colon cancer, gastric cancer, melanoma, head and neck cancer (HNC), pancreatic cancer, or a combination thereof.In some embodiments of the method, the head and neck cancer (HNC) is squamous cell carcinoma of the head and neck (SCCHN), unresectable HNC, squamous cell carcinoma of the head and neck (HNSCC), human papillomavirus (HPV)-positive head and neck cancer, metastatic HNC, metastatic HNC with three or more metastatic sites, occult primary metastatic squamous cell carcinoma, pharyngeal cancer, hypopharyngeal cancer, laryngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, oral cancer, sinus cancer, nasal cavity sinus cancer, nasal cavity cancer, salivary gland cancer, thyroid cancer, recurrent HNC, refractory HNC, HNC resistant to PD-1 therapy, HNC resistant to PD-L1 therapy, HNC resistant to immunotherapy, HNC resistant to T-cell therapy, HNC resistant to CAR-T therapy, HNC resistant to NK-cell therapy, or a combination thereof. In some embodiments of the method, the initiator is configured to include a hydrophobic electrophilic molecule. In some embodiments of the method, the initiator is configured to comprise a hydrocarbon. In some embodiments of the method, the hydrocarbon is configured to contain 1 to 22 carbon atoms, and the hydrocarbon may be saturated or unsaturated. In some embodiments of the method, the initiator is configured to comprise an aliphatic hydrocarbon, an aromatic hydrocarbon, or a combination of both. In some embodiments of the method, the initiator is configured to comprise a halide functional group. In some embodiments of the method, the initiator is configured to comprise an alkyl halide, an aralkyl halide, an acyl halide, or a combination thereof. In some embodiments of the method, the initiator is configured to include methyl iodine, methyl bromide, methyl chloride, ethyl iodine, ethyl bromide, ethyl chloride, 1-iodopropane, 1-bromopropane, 1-chloropropane, 1-iodobutane, 1-bromobutane, 1-chlorobutane, 1-iopentane, 1-bromopentane, 1-chloropentane, 1-iohexane, 1-bromohexane, 1-chlorohexane, 1-iododecane, 1-bromododecane, 1-chlorododecane, 1-iooctadecane, 1-bromooctadecane, 1-chlorooctadecane, benzyl iodine, benzyl bromide, benzyl chloride, allyl bromide, acyl iodine, acyl bromide, acyl chloride, benzoyl bromide, benzoyl chloride, or combinations thereof. In some embodiments of the method, the initiator is configured to contain a p-toluenesulfonyl group. In some embodiments of the method, the taxane is configured to associate with the at least one first end group. In some embodiments of the method, the polyoxazoline is configured to include poly(2-oxazoline), poly(2-substituted oxazoline), or combinations thereof. In some embodiments of the method, the polyoxazoline is configured to include poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-butyloxazoline), or combinations thereof. In some embodiments of the method, the taxane is configured to include paclitaxel, docetaxel, or combinations thereof. In some embodiments of the method, the aggregate comprises a polyoxazoline to taxane ratio of 7:1.In some embodiments of the method, the first pharmaceutical composition is administered to the subject via parenteral administration, the parenteral administration being selected from intravenous (IV) injection, intradermal injection, subcutaneous injection, or a combination thereof. In some embodiments of the method, the method further includes the step of administering an effective dose of a second pharmaceutical composition to the subject simultaneously or sequentially with the first pharmaceutical composition, the second pharmaceutical composition comprising carboplatin, cisplatin, an EGFR-binding monoclonal antibody or fragment thereof, a bispecific antibody or fragment thereof having at least one EGFR-binding site, a trivalent antibody or fragment thereof having at least one EGFR-binding site, a small molecule drug binding EGFR, or a combination thereof. In some embodiments of the method, the first and second pharmaceutical compositions are formulated as a single-dose unit comprising the first and second pharmaceutical compositions, the single-dose unit being selected from injectable packages containing the first and second pharmaceutical compositions separately in a co-package, premixed injectable packages containing a mixture of the first and second pharmaceutical compositions, or a combination thereof. In some embodiments of the method, the EGFR-binding monoclonal antibody or fragment thereof is cetuximab. In some embodiments of the method, the effective dose of the first pharmaceutical composition comprises 15 mg / m². 2 Up to 500 mg / m 2 The taxane is within the range described above, and the effective dose of the second pharmaceutical composition comprises 50 mg / m². 2 Up to 900 mg / m 2 The cetuximab is within the range described. In some embodiments of the method, the effective dose of the first pharmaceutical composition and the effective dose of the second pharmaceutical composition are administered to the subject at least once on day 1, day 8, day 15, day 28, or a combination thereof during a treatment cycle. In some embodiments of the method, the aggregate is configured to further comprise a targeting portion. In some embodiments of the method, the targeting portion is configured to comprise an antibody or a fragment thereof, its antigen-binding portion, an antigen, a cell surface receptor, a cytoplasmic receptor, a cell receptor ligand, or a lectin ligand.

[0009] In some cases, the methods disclosed herein relate to a method for treating cancer in subjects with corresponding needs, wherein said cancer may be breast cancer, triple-negative breast cancer, ovarian cancer, lung cancer, NSCLC (non-small cell lung cancer), colon cancer, gastric cancer, melanoma, head and neck cancer (HNC), pancreatic cancer, or a combination thereof. Head and neck cancer (HNC) can be squamous cell carcinoma of the head and neck (SCCHN), unresectable HNC, squamous cell carcinoma of the head and neck (HNSCC), human papillomavirus (HPV) positive head and neck cancer, metastatic HNC, metastatic HNC with 3 or more metastatic sites, occult primary metastatic squamous cell carcinoma, pharyngeal cancer, hypopharyngeal cancer, laryngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, oral cancer, sinus cancer, nasal cavity sinus cancer, nasal cavity cancer, salivary gland cancer, thyroid cancer, recurrent HNC, refractory HNC, HNC resistant to PD-1 therapy, HNC resistant to PD-L1 therapy, HNC resistant to immunotherapy, HNC resistant to T-cell therapy, HNC resistant to CAR-T therapy, HNC resistant to NK-cell therapy, or combinations thereof.

[0010] This disclosure provides a pharmaceutical composition comprising a first pharmaceutical composition comprising an aggregate comprising: (a) a polyoxazoline comprising at least one first end group modified with a hydrophobic portion, wherein the polyoxazoline further comprises a linear portion, a branched portion, or both, the branched portion comprising an asymmetricly branched polymer; and the polyoxazoline comprising a monomer to initiator molar ratio of 60:1; and (b) taxane, wherein the aggregate comprises a polyoxazoline to taxane weight ratio of 6:1 to 8:1; and a size of 70 nm to 90 nm. nm; and the polyoxazoline comprises a second end group containing a functional group modified with ethylenediamine (EDA) or an EDA derivative, the ratio of the second end group to EDA being 1:10; and a second pharmaceutical composition comprising carboplatin, cisplatin, a monoclonal antibody or fragment thereof binding to EGFR, a bispecific antibody or fragment thereof having at least one binding site for EGFR, a trivalent antibody or fragment thereof having at least one binding site for EGFR, a small molecule drug or combination thereof binding to EGFR; wherein the pharmaceutical composition is formulated as a single-dose unit comprising the first pharmaceutical composition and the second pharmaceutical composition containing the taxane, the single-dose unit being selected from injectable packages of the first pharmaceutical composition and the second pharmaceutical composition individually contained in a co-package, premixed injectable packages comprising a mixture of the first pharmaceutical composition and the second pharmaceutical composition, or combinations thereof. In some embodiments of the pharmaceutical composition, the monoclonal antibody or fragment thereof binding to EGFR is cetuximab. In some embodiments of the pharmaceutical composition, the pharmaceutical composition further comprises a salt component comprising sodium bicarbonate, and wherein the pH of the pharmaceutical composition is in the range of 7.1 to 10.

[0011] This disclosure also provides the use of the first and second pharmaceutical compositions in the manufacture of a medicament for treating head and neck cancer in a subject with corresponding need, wherein the first pharmaceutical composition comprises an aggregate comprising: (a) a polyoxazoline comprising at least one first end group modified by a hydrophobic portion, wherein the polyoxazoline further comprises a linear portion, a branched portion, or both, the branched portion comprising an asymmetricly branched polymer; and the polyoxazoline comprises a monomer to initiator molar ratio of 60:1, and (b) taxane, wherein the aggregate comprises a polyoxazoline to taxane weight ratio of 6:1 to 8:1; and a size of 70 nm to 90 nm. nm; and the polyoxazoline comprises a second end group containing a functional group modified with ethylenediamine (EDA) or an EDA derivative, wherein the ratio of the second end group to EDA is 1:10; and wherein the second pharmaceutical composition comprises carboplatin, cisplatin, a monoclonal antibody or fragment thereof that binds EGFR, a bispecific antibody or fragment thereof having at least one binding site for EGFR, a trivalent antibody or fragment thereof having at least one binding site for EGFR, a small molecule drug that binds EGFR, or a combination thereof; wherein the drug is formulated as a single-dose unit comprising the first pharmaceutical composition and the second pharmaceutical composition, the single-dose unit being selected from injectable packages containing the first pharmaceutical composition and the second pharmaceutical composition separately in a co-package, premixed injectable packages comprising a mixture of the first pharmaceutical composition and the second pharmaceutical composition, or a combination thereof. In some embodiments of the use of the first pharmaceutical composition and the second pharmaceutical composition in the manufacture of a drug, the monoclonal antibody or fragment thereof that binds EGFR is cetuximab. In some embodiments of the use of the first and second pharmaceutical compositions in the manufacture of a medicament, an effective dose of the first pharmaceutical composition and an effective dose of the second pharmaceutical composition are administered to the subject at least once on day 1, day 8, day 15, day 28, or a combination thereof during a treatment cycle.

[0012] Additional features and advantages of this disclosure are described in the following detailed description and accompanying drawings, and will be apparent from the following detailed description and accompanying drawings.

[0013] Symmetrically branched polymers (SBP) A class of polymers called dendrimers, including starburst dendrimers (or dense star-shaped polymers) and combburst dendrigrafts (or supercomb-shaped branched polymers), have recently been developed and studied for various industrial applications. These polymers typically possess: (a) a well-defined core molecule, (b) at least two concentric dendritic layers (generations) with symmetrical (equal length) branches and branch junctions, and (c) outer surface groups, such as branched and dendritic polymers based on polyamide-amine (PAMAM). Other examples include polyethyleneimine (PEI) dendrimers, polypropyleneimine (PPI) dendrimers, Frechet-type polyether and polyester dendrimers, core-shell tectodendrimers, and others.

[0014] Comb-like burst dendritic grafts are constructed using a stepwise synthetic approach with a core molecule and concentric layers exhibiting symmetrical branching. Unlike dendritic polymers, comb-like burst dendritic grafts or polymers are generated from monodisperse linear polymer structural units. Furthermore, the branching patterns differ from those of dendritic polymers. For example, comb-like burst dendritic grafts form branch junctions (chain branching) along the polymer backbone, while starburst dendritic polymers typically branch at the ends (terminal branching). Due to the living polymerization techniques used, the molecular weight distribution (Mw / Mn) of these polymers (core and branches) is typically narrow. Therefore, comb-like burst dendritic grafts produced via graft-regrafting processes are well-defined, where the Mw / Mn ratio is typically close to 1.

[0015] SBPs, such as dendritic polymers, are primarily produced through divergent or metamerization methods via repeated protection and deprotection processes. Because dendritic polymers utilize small molecules as core and branching structural units, their molecular weight distribution is typically limited. At lower generations, dendritic polymers with a single molecular weight are usually obtained. While dendritic polymers generally utilize small monomers as structural units, dendritic grafts use linear polymers as structural units.

[0016] In addition to dendritic polymers and dendritic grafts, other SBPs include symmetrical star or comb polymers, such as symmetrical star or comb polyethylene oxide (PEO), polyethylene glycol (PEG), polyoxazoline (POX), polymethyloxazoline (PMOX), polyethyloxazoline (PEOX), polystyrene, polymethyl methacrylate, polydimethylsiloxane, or any combination thereof.

[0017] Asymmetric branched polymers (ABP) Unlike SBPs, asymmetricly branched polymers (ABPs), particularly asymmetrically branched dendritic polymers or regular ABPs (reg-ABPs), typically have a core, controlled and well-defined asymmetric (unequal length) branches, and asymmetric branch junctions. Random ABPs, on the other hand, have: (a) no core, (b) functional groups both externally and internally, (c) random / variable branch lengths and patterns (i.e., terminal and chain branching), and (d) a non-uniformly distributed internal void space.

[0018] The synthesis and mechanism of ran-ABP (e.g., those made from PEI) have been previously reported, as have those made from POX, namely poly(2-methyloxazoline) and poly(2-ethyloxazoline). The synthesis of ran-ABP can generally involve either a one-pot divergent method or a one-pot polymeric method.

[0019] homopolymer Homopolymers can refer to polymers or polymer backbones containing the same repeating units, i.e., homopolymers are generated from the same monomers (e.g., linear PEI polymers, linear POX polymers, dendritic PEI polymers, polyamide-amine (PAA) dendritic polymers, or POX dendritic grafts and random branched polymers). Monomers can be simple compounds or complexes or aggregates of compounds, where the aggregate or complex is the repeating unit in the homopolymer. Thus, if an aggregate contains three compounds A, B, and C, the complex can be described as ABC. For the purposes of this disclosure, a polymer containing (ABC)-(ABC)-(ABC) is a homopolymer because the backbone contains a single simple or complex monomer. Homopolymers can be linear or branched. Thus, in the case of randomly branched PEI, although branches of varying lengths exist and the branches appear randomly, for the purposes of this disclosure, the molecule is a homopolymer because the branched polymer contains a single monomer, ethyleneimine, or aziridine repeating unit. Furthermore, one or more of the monomeric or complex monomeric components can be modified, substituted, derivatized, etc.; for example, modified to carry functional groups. Brief description of the attached diagram The following description of the accompanying drawings and corresponding figures is a non-limiting example illustrating various embodiments of the present disclosure.

[0021] Figure 1A - Figure 1D The structure of SBP is shown, for example, a dendritic polymer ( Figure 1A ), dendritic grafts ( Figure 1B ), comb-shaped polymers ( Figure 1C ) and star polymers ( Figure 1DAll polymers have a spherical or linear core.

[0022] Figure 2A - Figure 2B It shows a branch with 4 branches ( Figure 2A ) and 8 branches ( Figure 2B The chemical structure of symmetrically branched PPI dendritic polymers.

[0023] Figure 3 The chemical modification reactions of symmetrically branched PPI dendritic polymers are depicted, where the numbers 8, 16, 32, etc. indicate the number of reactive groups on the surface of the dendritic polymer.

[0024] Figure 4A - Figure 4B This illustrates a random ABP with asymmetric branch junctions and patterns. Figure 4A ) and rule ABP ( Figure 4B The structure of ).

[0025] Figure 5 The chemical structures of randomly asymmetric branched PEI homopolymers were described.

[0026] Figure 6A - Figure 6B The synthesis scheme is shown, in which Figure 6A It is a chemical modification reaction of randomly asymmetric branched PEI homopolymers, and Figure 6B This is a one-pot synthesis of randomly branched poly(2-ethyloxazoline) with hydrophobic modification of primary amino groups at the polymer focal points. The initiator / surface group (I) is a brominated hydrocarbon, and the reaction opens the oxazoline ring.

[0027] Figure 7 A﹣ Figure 7 B shows the SBP( Figure 7 A) and ABP Figure 7 B) The structure of the drug loaded in the surface domain or region or at the surface domain or region, where R represents the surface group and the solid circle represents the drug of interest.

[0028] Figure 8 Representative structures of composite-based nanoparticles containing both drug molecules and branched polymers are shown.

[0029] Figure 9 A﹣ Figure 9 B shows that in SBP ( Figure 9 A) and ABP Figure 9 B) is a representative structure of an insoluble or poorly water-soluble drug (represented by a solid circle) loaded at a hydrophobic surface group (represented by a wavy line).

[0030] Figure 10A- Figure 10B The representative structure of drug-containing nanoparticles is shown. Figure 10A , Figure 10B The drug-containing nanoparticles also carry at least one targeting group or portion, denoted as “Y”, such as an antibody.

[0031] Figure 11 This is a graph comparing the sizes of polymer-only aggregates and polymer-drug aggregates in saline at polymer concentrations of 25 mg / mL and drug concentrations of 5 mg / mL. The polymer is hydrophobically modified random branched PEOX, and the drug is paclitaxel.

[0032] Figure 12 This is a graph comparing the sizes of polymer-only aggregates and polymer-drug aggregates in saline at polymer concentrations of 2.5 mg / mL and drug concentrations of 0.5 mg / mL. The polymer is hydrophobically modified random branched PEOX, and the drug is paclitaxel.

[0033] Figure 13 This is a graph comparing the sizes of polymer-only aggregates and polymer-drug aggregates in saline at polymer concentrations of 250 μg / mL and drug concentrations of 50 μg / mL. The polymer is hydrophobically modified random branched PEOX, and the drug is paclitaxel.

[0034] Figure 14 This is a graph comparing the sizes of polymer-only aggregates and polymer-drug aggregates in saline at polymer concentrations of 25 μg / mL and drug concentrations of 5 μg / mL. The polymer is hydrophobically modified random branched PEOX, and the drug is paclitaxel.

[0035] Figure 15 This is a graph depicting cell survival rates after exposure to three taxane formulations.

[0036] Figure 16 This is a graph depicting the cytotoxicity of A549 lung cancer cells after exposure to three different taxane formulations.

[0037] Figure 17 This is a graph depicting the cytotoxicity of MDA-MB-231 triple-negative breast cancer cells after exposure to three different taxane formulations.

[0038] Figure 18 This is a graph depicting the toxicity of OV-90 ovarian cancer cells after exposure to three different taxane formulations.

[0039] Figure 19 This is a graph depicting the pharmacokinetic (PK) profiles of three different taxane formulations, showing changes in plasma concentration over time.

[0040] Figure 20 This is a graph depicting the tumor volume of A549 lung cancer in mouse xenograft models treated with two control treatments and exposed to three different taxane formulations.

[0041] Figure 21 Representative images of resected lung cancer cell tumors growing as xenografts in mice are shown, along with mice treated with two control groups and two forms of taxane.

[0042] Figure 22 This figure depicts the effects of two negative controls and three taxane preparations on ovarian cancer tumor size in a mouse xenograft model.

[0043] Figure 23 Representative images of excised ovarian cancer cell tumors growing as xenografts in mice are shown, along with mice treated with two controls and three forms of taxane.

[0044] Figure 24A - Figure 24B Representative images showing tumor size reduction after treatment with FID-007 are shown. Figure 24A The size of the tumor before treatment was shown, and Figure 24B The size of the tumor after treatment is shown.

[0045] Detailed Explanation By reading the following detailed description, those skilled in the art will more readily understand the features and advantages of this disclosure. It should be understood that certain features of this disclosure described in the context of different embodiments above and below may also be provided in combination in a single embodiment. Conversely, for the sake of brevity, various features of this disclosure described in the context of a single embodiment may also be provided individually or in any combination or sub-combination. Furthermore, unless the context clearly indicates otherwise, singular references may also include plural references (e.g., "a" and "an" may refer to one or more).

[0046] Unless otherwise expressly indicated, the use of numerical values ​​within the various ranges specified in this application is to indicate approximations, as the minimum and maximum values ​​within the range are preceded by the word "approximately". In this way, slightly higher and lower than the range can achieve substantially the same result as values ​​within the range. Furthermore, the disclosure of ranges is intended to be a continuous range including every value between the minimum and maximum values, and including both the minimum and maximum reference values. For the purposes of this disclosure, words such as "approximately" and "substantially" are defined as a range of values ​​no greater than 10% of the stated value or number.

[0047] In this disclosure, drug solubility is defined as the number of solvent parts required to dissolve one part of the drug, <30 (soluble), 30-100 (poorly soluble), and >100 (insoluble). When water is used as a solvent, taxanes such as paclitaxel and its derivatives are water-insoluble or poorly soluble in water.

[0048] "Homopolymers" are as described above.

[0049] As used herein, the term "combined positive score" (CPS) refers to an algorithm that includes both tumor and immune cells to determine the expression of programmed death-ligand 1 (PD-L1) protein in tumor tissue. In some instances, CPS = (number of PD-L1-stained cells (tumor cells, lymphocytes, macrophages)) / (total number of viable tumor cells) × 100. Some instances may include FDA-approved, clinically validated CPS diagnostic cutoff values ​​used to determine the expression of PD-L1-stained cells in KEYTRUDA. ® The six tumor indications in the patients treated were: gastric or gastroesophageal junction (GC / GEJ) adenocarcinoma (CPS≥1), cervical cancer (CPS≥1), urothelial carcinoma (CPS≥10), head and neck squamous cell carcinoma (HNSCC) (CPS≥1), esophageal squamous cell carcinoma (ESCC) (CPS≥10), and triple-negative breast cancer (TNBC) (CPS≥10).

[0050] Drugs of interest The drug of interest described herein is taxane, and includes paclitaxel and other taxane derivatives such as docetaxel. Paclitaxel is water-insoluble and has well-defined performance characteristics, such as a low maximum tolerated dose (MTD), PK profile, and limited potency in treating various types of cancer. This disclosure covers the use of ABP, as previously described, in improving those performance characteristics.

[0051] Nanocomposite materials, nanoparticles or nanoaggregates Nanocomposites are physical mixtures of two or more materials or components (e.g., polymers and taxanes). In this disclosure, such mixtures may contain different nanoscale phases or domains, solid or liquid, formed between taxane and branched homopolymer molecules. Nanocomposites may comprise a combination of a bulk matrix (e.g., branched homopolymers and taxanes) and nanoscale two-dimensional phases that may exhibit different properties due to differences in structure and chemistry (e.g., domains formed by surface groups of taxanes and branched polymers, and domains formed within the branched polymer). Because the solubility of domains / phases may differ, when dissolving a nanocomposite in an aqueous solution, one phase may dissolve faster than another or other phases, leading to gradual decomposition of the composite aggregates, resulting in the hierarchical and controlled release of the composite components, and optionally, the reformation of one or more components into new forms, such as new aggregates. The terms “nanocomposites,” “nanoparticles,” and “nanoclusters” are equivalent and used interchangeably herein.

[0052] The aggregates described in this disclosure range in size from about 10 nm to about 500 nm in diameter, and from about 30 nm to about 300 nm in diameter. The aggregates may exhibit size-related properties that are significantly different from those observed in particulate or bulk materials.

[0053] SBP is described in Figure 1A - Figure 1D In this model, there is symmetrical branching, where all homopolymers of interest have a core and exhibit symmetrical branch junctions consisting of terminal or chain branches throughout the homopolymer. Functional groups are primarily located on the exterior.

[0054] Modified SBPs can be obtained, for example, by chemically linking functional groups on, for example, symmetrically branched PAMAM or PPI dendritic polymers (commercially available from Aldrich), polyether dendritic polymers, polyester dendritic polymers, comb-branched / star-branched polymers (e.g. those containing PEO, PEG, PMOX, or PEOX), polystyrene, and comb-branched dendritic grafts (e.g. those containing PEOX, PMOX, or PEI).

[0055] Use commercially available reagents (e.g., various generations of PPI dendritic polymers, such as dendritic polymer-4). Figure 2A ) and dendritic polymer -8( Figure 2B Synthetic procedures for preparing such SBP / dendritic polymers are known, or many SBPs are commercially available. The synthesis of comb-branched polymers and comb-bursting polymers is also known. Symmetrically branched PPI dendritic polymers can be prepared, for example, via... Figure 3The illustrated reactions are used for chemical modification. The numbers 8, 16, 32, 64, or 128 indicate the number of reactive groups on the surface of the dendritic polymer.

[0056] SBP’s high branching density makes the polymer molecularly compact with well-defined internal void spaces, making it suitable as a carrier for embedding or encapsulating taxanes.

[0057] Surface modification can enhance the properties and uses of the resulting modified SBPs. For example, with appropriate modification, water-insoluble SBPs can be made water-soluble, while SBPs with high charge density can be modified to carry very low or no charge on or at the polymer surface. On the other hand, water-soluble SBPs can be modified with hydrophobic surface groups to enhance their ability to dissolve water-insoluble or poorly soluble drugs on their surface. Modification can occur at any site on the polymer, such as at the ends, branches, main chain residues, etc.

[0058] In one embodiment of this disclosure, the SBP (e.g., such as symmetrically branched PEI dendritic polymers, PPI dendritic polymers, PAMAM dendritic polymers, or symmetrically branched PEI dendritic grafts) can be modified with different kinds of, for example, primary amine groups by, for example, Michael addition or by adding acrylate to the amine groups of the homopolymer. Thus, for example, methyl acrylate can be introduced onto the primary and / or secondary amine groups of PEI, PPI, and polylysine (PLL) homopolymers via Michael addition. The ester groups can then be further derivatized, for example, via amidation. Thus, for example, such amidation with, for example, ethylenediamine (EDA) can produce the addition of amine groups at the ends of the newly formed branches. Other modifications to the homopolymer can be made using known chemistry, for example, as provided in "Dendrimers and Other Dendritic Polymers," edited by Frechet & Tomalia, John Wiley & Sons, Ltd., 2001. Derivatives of EDA may also be used, and include any molecular entity containing reactive EDA, substituted EDA, or other members of the polyethyleneamine family, such as diethylenetriamine, triethylenetetramine, tetraethylenepentamine, pentaethylenehexamine, etc., including polyethyleneamine, tetramethylethylenediamine, etc.

[0059] In some embodiments, the modification may include a portion that contributes to or enhances the hydrophobicity of the polymer or a portion of the polymer. For example, hydrophobic functional groups, such as aliphatic chains (e.g., hydrocarbon chains containing one to about 22 carbons, whether saturated or unsaturated, linear, cyclic or branched), aromatic structures (e.g., containing one or more aromatic rings, which may be fused), or combinations thereof, may be used as modifiers and added to the polymer, as taught herein and practiced as provided herein in chemistry.

[0060] Upon this addition, modified SBPs are formed, such as modified PEI, PPI, PAMAM dendrimers, or PEI dendritic grafts. As extensions of SBPs such as PPI and PEI, the resulting modified SBPs are also symmetrically branched. Depending on the solvent environment (i.e., pH or polarity), the surface functional groups can carry different charges and / or charge densities, and / or hydrophobic groups. The molecular shape and the position of the surface functional groups (i.e., surface functional group foldback) can then be further tuned based on those characteristic properties.

[0061] In another embodiment of this disclosure, the modified SBP can be generated using any of a variety of synthetic schemes, for example, synthetic schemes known to be suitable for reacting at appropriate sites on the homopolymer. Furthermore, any of a variety of reagents can be used in the chosen synthetic scheme to generate any of a variety of modifications or additions to the homopolymer backbone. Thus, for example, in the case of a Michael addition reaction of an amine described above, such as in the alkylation stage, addition of any of a variety of substituents can be used, using, for example, any of a variety of acrylate reagents, such as acrylates containing hydrocarbon substituents, said hydrocarbon substituents comprising, for example, saturated or unsaturated hydrocarbons of about 1 to 22 carbons, which can be substituted, aliphatic, aromatic, cyclic, saturated at one or more bonds, or combinations thereof. Therefore, suitable reactants include methyl acrylate, ethyl acrylate, propyl acrylate, butyl acrylate, pentyl acrylate, hexyl acrylate, heptyl acrylate, octyl acrylate, nonyl acrylate, decyl acrylate, undecyl acrylate, dodecyl acrylate, etc., and mixtures thereof. Similarly, in the amidation stage of the examples illustrated above, any of a variety of amines can be used. For example, EDA, monoethanolamine, tris(hydroxymethyl)aminomethane, alkylamine, allylamine, or any amino-modified polymer can be used, including those containing: PEG, PEO, perfluoropolymers, polystyrene, polyethylene, polydimethylsiloxane, polyacrylate, polymethyl methacrylate, etc., and mixtures thereof.

[0062] This synthetic strategy not only allows for symmetrical molecular growth (where more branches with different chemical compositions can be introduced), but also allows for the addition of multiple functional groups to the exterior of the structure. Precursor homopolymers can be continuously modified using the same or different synthetic processes until the desired SBP with appropriate molecular weight and functional groups is obtained. Furthermore, the hydrophobic and hydrophilic properties of this polymer, as well as its charge density, can be tailored to specific application needs using appropriate monomers for constructing the homopolymer and suitable modification reactions.

[0063] In another embodiment of this disclosure, if a divergent synthesis procedure is used, the chain ends of the symmetrical star-branched homopolymer or comb-branched homopolymer can be modified with another small molecule or polymer to generate various functional groups at the homopolymer chain ends. The symmetrical star-branched homopolymer or comb-branched homopolymer includes, for example, poly(2-substituted oxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), and poly(2-butyloxazoline), poly(2-substituted oxazoline), PEI, PEO / diol, polyvinylpyrrolidone (PVP), polyphosphate, polyvinyl alcohol (PVA), or polystyrene. The functional groups include primary, secondary, or tertiary amines, carboxylic esters, hydroxyl groups, aliphatic (e.g., hydrocarbon chains), aromatic groups, fluoroalkyl groups, aryl groups, PEG, PEO, acetate groups, amide groups, and / or ester groups. Alternatively, various initiators can be used so that if a polymerization method is used, the same type of functional groups can be introduced at the end of the chain.

[0064] Initiators can be hydrophobic, electrophilic molecules, including hydrocarbons, aliphatic hydrocarbons, aromatic hydrocarbons, or combinations thereof, and halogen functional groups, such as alkyl halides, aralkyl halides, acyl halides, or combinations thereof. Examples of such compounds are monofunctional initiators, such as hydrocarbons containing one to about 22 carbons with saturated or unsaturated bonds, such as methyl iodide / methyl bromide / methyl chloride, ethyl iodide / ethyl bromide / ethyl chloride, 1-iodobutane / 1-bromobutane / 1-chlorobutane, 1-iodohexane / 1-bromohexane / 1-chlorohexane, 1-iododecane / 1-bromododecane / 1-chlorododecane, 1-iodooctadecane / 1-bromooctadecane / 1-chlorooctadecane, benzyl iodide / benzyl bromide / benzyl chloride, etc. Other initiators include allyl bromide / allyl chloride. Acyl halides, such as acyl bromide / acyl chloride, benzoyl bromide / benzoyl chloride, and compounds containing p-toluenesulfonyl groups, such as p-toluenesulfonic acid, methyl p-toluenesulfonate, and other p-toluenesulfonate esters, can also be used. Any one or more initiators can be used in combination.

[0065] During polymerization, an initiator can be used to start the polymerization process. When used, various monomer-to-initiator molar ratios can be used to obtain specific polymers. Specific polymers can have different properties, such as molecular size. Therefore, suitable monomer-to-initiator molar ratios can range from 20:1 to 80:1, for example 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, or 75:1, including 21:1, 22:1, 23:1, 24:1, 26:1, 27:1, 28:1, 29:1, 31:1, 32:1, 33:1, 34:1, 36:1, 37:1, 38:1, 39:1, 41... :1, 42:1, 43:1, 44:1, 46:1, 47:1, 48:1, 49:1, 51:1, 52:1, 53:1, 54:1, 56:1, 57:1, 58:1, 59:1, 61:1, 62:1, 63:1, 64:1, 66:1, 67:1, 68:1, 69:1, 71:1, 72:1, 73:1, 74:1, 76:1, 77:1, 78:1, 79:1, and any other ratio within this range.

[0066] ABP is described in Figure 4A and Figure 4B In homopolymers, asymmetric branching is present, with some polymers of interest lacking a core and exhibiting asymmetric branch junctions consisting of both chain and terminal branches throughout the homopolymer. Linking groups are typically present both externally and internally. However, when larger functional groups (e.g., large hydrophobic or hydrophilic groups) are used, these functional groups can often preferentially and possibly necessarily attach to the exterior of the ABP, perhaps due to steric effects. Therefore, such surface MBPs can be used to dissolve insoluble or poorly soluble drugs or to form aggregates with insoluble or poorly soluble drugs.

[0067] Modified ABPs can be, for example, chemically linked to regular ABPs such as polylysine (e.g., branched PLL) or random ABPs such as PEI (available from Aldrich, Polysciences, or BASF under the trade name LUPOSAL). TM Commercially available ABPs (ABPs) or functional groups on polyoxazoline can be obtained, and the regular, random, or polyoxazoline ABPs can be prepared according to known procedures. Other ABPs may include, but are not limited to, polyacrylamide, polyphosphate, PVP, PVA, etc.

[0068] Such modified ABPs can be prepared using a variety of known starting materials. These monomers and polymers are commercially available in large quantities at a reasonable cost. For example, one such precursor monomer that can be used to synthesize the homopolymer of interest is PEI. The synthesis of randomly branched PEI is known. PEIs of various molecular weights are available from sources such as Aldrich, Polysciences, and BASF (under the trade name LUPOSAL). TM Randomly branched PEIs are commercially available from various sources. They are primarily produced via cationic ring-opening polymerization of cyclostrained cyclic imine monomers such as aziridine (ethyleneimine) and aziridine (propyleneimine), with Lewis or Brønsted acids acting as initiators. Since many methods are essentially one-pot processes, large quantities of random ABPs can be readily produced. Randomly branched poly(2-substituted oxazoline) polymers can be prepared using known procedures.

[0069] The synthetic processes used to prepare ABP typically generate various branching junctions within the macromolecule. In other words, a mixture of terminal junctions and chain branching junctions is distributed throughout the molecular structure. Random ABPs can exhibit lower branching density and a more open molecular structure compared to dendritic polymers and dendritic grafts. Although the branching pattern is random, the average ratio of primary, secondary, and tertiary amine groups can be relatively consistent with an approximate 1:2:1 ratio.

[0070] The presence of branching junctions allows random ABPs (such as asymmetrically branched PEI) to form macromolecules with possible spherical, oval, or similar configurations. Within the spherical structure, there are bags of various sizes formed by imperfect branching junctions within the macromolecule. Unlike dendritic polymers and dendritic grafts, where the internal bags are always located around the central core of the molecule, the bags in random ABPs are unevenly distributed throughout the molecule. Therefore, random ABPs possess both external functional groups and unevenly distributed internal functional groups, which can further react with various molecules to form new macromolecular structures—the modified random ABPs of interest.

[0071] Despite having a core, the functional groups of regular ABPs are also distributed on both the exterior and interior, much like random ABPs. One such homopolymer is PLL. Such homopolymers can also be modified in a manner similar to that of random ABPs, as taught herein and as known in the art.

[0072] In embodiments of this disclosure, ABPs (e.g., randomly asymmetrically branched PEI or regularly asymmetrically branched PLLs) are modified with different kinds of primary amine groups by, for example, Michael addition or by adding acrylates to the amines of the polymer. Thus, for example, by Michael addition, methyl acrylate or other acrylates as provided herein can be introduced onto the primary and / or secondary amine groups of, for example, PEI and PLL homopolymers. The ester groups can then be further derivatized, for example, by amidation. Thus, for example, such an amidation reaction with, for example, EDA, can produce an addition of amine groups at the ends of newly formed branches. Other modifications to the polymer can be made using known chemistry. Examples of randomly asymmetrically branched PEI homopolymers are shown in... Figure 5 middle.

[0073] Upon this addition, modified ABPs are formed, such as modified PEI or PLL homopolymers. As extensions of ABPs like PEI and PLL, the resulting modified ABPs are also asymmetrically branched. Depending on the solvent environment (i.e., pH or polarity), the surface functional groups can carry different charges and charge densities. The molecular shape and functional group positions (i.e., functional group foldback) can then be further tuned based on those characteristic properties.

[0074] In another embodiment, any of a variety of synthetic schemes can be used to produce the modified ABP, for example, the synthetic schemes are known to be suitable for reacting at suitable sites on the homopolymer. Furthermore, any of a variety of reagents can be used in the chosen synthetic scheme to produce any of a variety of modifications or additions to the polymer backbone. Thus, for example, in the case of the Michael addition reaction of the amine described above, addition of any of a variety of substituents can be used in the alkylation stage, as provided above, for example using acrylates, which can include saturated or unsaturated hydrocarbons, such as hydrocarbons containing 1 to about 22 carbons, which can be aliphatic, branched, saturated, aromatic, cyclic, or combinations thereof. Suitable reactants include methyl acrylate, ethyl acrylate, propyl acrylate, butyl acrylate, pentyl acrylate, hexyl acrylate, heptyl acrylate, octyl acrylate, nonyl acrylate, decyl acrylate, undecyl acrylate, dodecyl acrylate, etc., and mixtures thereof. Similarly, in the amidation stage of the examples illustrated above, any of a variety of amines can be used in the methods provided herein and known in the art. For example, EDA, monoethanolamine, tris(hydroxymethyl)aminomethane, alkylamine, allylamine, or any amino-modified polymers, including PEG, perfluoropolymers, polystyrene, polyethylene, polydimethylsiloxane, polyacrylate, polymethyl methacrylate, etc., and mixtures thereof, can be used. Furthermore, aliphatic polymers (e.g., from C1 to about C4) can also be used. 22The connection of hydrophobic groups (including hydrocarbon groups), aromatic groups, polyethylene polymers, polystyrene polymers, perfluoropolymers, polydimethylsiloxane, polyacrylates, and polymethyl methacrylate, as well as hydrophilic groups (including OH groups) and hydrophilic groups of hydrophilic polymers such as PEOX, PEG, and PEO) to modified ABPs can be achieved using, for example, epoxidation reactions, amidation reactions, Michael addition reactions, including the use of -SH or -NH2 groups reacting with maleimide, aldehyde / ketone-amine / hydrazide coupling reactions, iodine / iodoacetyl-SH coupling reactions, hydroxylamine-aldehyde / ketone coupling reactions, etc. This synthetic strategy not only allows for asymmetric growth of molecules (where more bags are introduced), but also allows for the addition of multiple functional groups both inside and outside the structure. Homopolymers can be further modified using the same or different synthetic processes until the desired ABP with appropriate molecular weight and functional groups is obtained. Furthermore, the hydrophobic and hydrophilic properties, as well as the charge density, of such homopolymers can be customized using appropriate monomers and suitable modification reactions to suit specific application needs.

[0075] In another embodiment of this disclosure, the focus of the random ABP (e.g., POX) (which merges from various reactive chain ends during copolymerization) can be terminated or reacted with another small molecule to generate various functional groups at the homopolymer chain ends, including primary, secondary, or tertiary amines, carboxylic esters, hydroxyl, alkyl, fluoroalkyl, aryl, PEG, acetate, amide, and / or ester groups. Alternatively, various initiators can also be used such that the same type of functional group can be introduced at the surface group at the start of polymerization during copolymerization.

[0076] Randomly branched poly(2-ethyloxazoline) with alkyl surfaces modified with primary amine groups at the focal points of branched polymers can be prepared using known procedures. For example, CH3(CH)2) 17 Br can be used as an initiator for the polymerization of 2-ethyloxazoline, generating randomly branched polymers via a cationic ring-opening process, followed by quenching with N-tert-butoxycarbonylpiperazine (N-Boc-piperazine) or EDA. Termination with a large excess of EDA allows the hydrophobically modified branched poly(2-ethyloxazoline) polymer to be functionalized at the focal point with a primary amine group. Figure 6A and Figure 6B Alternatively, the N-Boc-piperazine-terminated hydrophobically modified branched poly(2-ethyloxazoline) polymer can also be deprotected to generate free amino groups at the focal point. If not terminated, the polymer focal point can be hydrolyzed into, for example, hydroxyl groups when dissolved in water (e.g., containing, for example, 1 N Na2CO3).

[0077] While introducing primary amine groups into hydrophobically modified branched poly(2-oxazoline) homopolymers enhances drug solubility and generates taxane-induced aggregates, primary amine groups also allow for the attachment of various targeting groups to, for example, hydrophobically modified branched poly(2-oxazoline) polymers. Figure 10A and Figure 10B The targeting group is, for example, an antibody or fragment thereof, its antigen-binding portion, an antigen, or a member of a binding pair. Aggregates or nanoparticles containing such a targeting group and its modifications can provide targeting capability on aggregates containing taxanes and enable the taxanes to be released preferentially or only at the desired therapeutic site.

[0078] As taught herein, MBPs, such as hydrophobically modified homopolymers including both SBPs and ABPs, can be used to produce encapsulated polymers or nanocapsules, to dissolve water-insoluble or poorly soluble taxanes, or to form taxane-induced nanoparticles of water-insoluble or poorly soluble taxanes such as paclitaxel. Figure 7 A, Figure 7 B. Figure 8 , Figure 9 A and Figure 9 B). In an organic solvent environment, the hydrophilic or amphiphilic interior can be poly(2-oxazoline), poly(2-substituted oxazoline) (including poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), and poly(2-butyloxazoline), etc.), PEG, PEO, polyphosphonates, etc. The hydrophobic exterior can contain aliphatic hydrocarbons (e.g., from C1 to about C1). 22 Aromatic hydrocarbons, polyethylene polymers, polystyrene polymers, perfluoropolymers, polydimethylsiloxane, polyacrylates, polymethyl methacrylate, etc. In an aqueous environment, the situation is exactly the opposite. In drug-induced aggregates in an aqueous environment, drug molecules (e.g., taxanes) associate with the hydrophobic groups / domains of MBPs (…). Figure 9 A and Figure 9 B). The branching density of branched homopolymers (e.g., from low-generation star-shaped and comb-shaped homopolymers to high-generation dendritic polymers and dendritic grafts) and the coverage of hydrophobic surface groups (e.g., from 0% to 100% coverage) can significantly affect the solubility of the homopolymer, which in turn affects its ability to dissolve or adsorb / absorb taxanes. For example, increasing the branching density and the coverage of hydrophobic groups will make the homopolymer more compatible with taxanes.

[0079] In some cases, ABPs and SBPs having about 0.1% to about 30% or more by weight of a surface-hydrophobic component can effectively dissolve or disperse water-poor or water-insoluble taxanes, such as paclitaxel. Furthermore, the branched homopolymers used, such as POX, PEOX, PMOX, PEO / PEG, polyacrylamide, polyphosphate, PVP, and PVA, are soluble in both water and a variety of organic solvents, thereby promoting the formation of various taxane-containing nanoparticles or aggregates. The good water solubility and good hydrophobic drug miscibility in aqueous solutions, with or without other organic solvents, make such homopolymers suitable for enhancing the solubility of water-poor taxanes. For example, homopolymers of interest simplify the manufacturing process and reduce production costs by reducing formulation steps, processing time, and the need for complex and expensive equipment currently used in the pharmaceutical industry. If additional branching density is required, the SBP or ABP can first be modified with additional groups as described herein, and then, for example, attached with additional hydrophobic functional groups to enhance taxane solubility.

[0080] When hydrophobically modified SBPs or ABPs are mixed with water-insoluble or poorly soluble taxanes (e.g., paclitaxel), different physical aggregates are formed, with sizes different from those formed solely by polymers. Figure 11 - Figure 14 As the concentrations of homopolymer and taxane decrease, the size and distribution of the polymer / taxane aggregates become much more similar to those of polymer-only aggregates, indicating that taxane is released from the induced aggregates or nanoparticles. The broad size distribution of polymer-only aggregates, regardless of association with taxane, is similar to that observed in other structures containing lipids. On the other hand, the taxane-induced aggregates of interest exhibit a specific size distribution with a narrower distribution, i.e., producing unique aggregates of a certain size. As the concentration of taxane in the aggregate decreases, the concentration of homopolymer in the aggregate decreases, the aggregate concentration decreases, or any combination thereof, the aggregates of interest release paclitaxel, as demonstrated by a decrease in aggregate size and / or a wider aggregate size distribution. Due to taxane release, a wider distribution may result from a mixture of homopolymer-only aggregates and polymer / taxane aggregates of different sizes, until the only aggregates observed are those with the characteristics of homopolymer-only aggregates. In other words, taxane is gradually released after introduction into the host, for example, in the circulatory system. This mechanism is important for a variety of drug delivery applications, including intravenous (IV), oral, transdermal, ocular, and intramuscular administration, which may require delayed or sustained release profiles.

[0081] For simplicity, the term "polymer" used to describe aggregates, polymer-drug nanoparticles, polymer aggregates, or polymer-to-drug ratios, etc., includes SBP and ABP disclosed herein, and includes modified polyoxazolines, such as hydrocarbon-modified polyoxazolines, including those further modified with EDA or EDA derivatives disclosed herein.

[0082] Suitable polymer-taxane weight ratios are from 6:1 to 8:1, for example, 6.5:1, 7:1, or 7.5:1, including every and all of 6.1:1, 6.2:1, 6.3:1, 6.4:1, 6.5:1, 6.6:1, 6.7:1, 6.8:1, 6.9:1, 7.0:1, 7.1:1, 7.2:1, 7.3:1, 7.4:1, 7.5:1, 7.6:1, 7.7:1, 7.8:1, 7.9:1, and 8.0:1, as well as all ratios within this range. In some cases, the polymer may be polyoxazoline, and the suitable polyoxazoline to taxane weight ratio is from 6:1 to 8:1, for example, 6.5:1, 7:1 or 7.5:1, including every and all of 6.1:1, 6.2:1, 6.3:1, 6.4:1, 6.5:1, 6.6:1, 6.7:1, 6.8:1, 6.9:1, 7.0:1, 7.1:1, 7.2:1, 7.3:1, 7.4:1, 7.5:1, 7.6:1, 7.7:1, 7.8:1, 7.9:1, 8.0:1, and all ratios within this range.

[0083] The applicant unexpectedly discovered that the combination of the molar ratio of monomer to initiator in polymerization and the weight ratio of polyoxazoline to taxane in nanoparticles can affect the large-scale manufacturability of drug nanoparticles, nanoparticle size, and efficacy as a tumor-reducing therapy. As an example, using a polymer synthesized at a monomer:initiator molar ratio of 100:1, taxane-induced aggregates prepared at a polymer:taxane weight ratio of 5:1 produced large nanoparticles, for example, in the 120 nm–140 nm range before lyophilization. When mass-produced, these large nanoparticles were difficult to pass through a 0.22 μm filter (required for the sterilization step in injections).

[0084] In contrast, when the polymer synthesized using a monomer:initiator molar ratio of 60:1 is mixed with taxane at a polymer:taxane weight ratio of 7:1, the resulting nanoparticles can have a size of 70 nm–100 nm before lyophilization, which allows the particles to pass through a 0.22 μm filter with almost no difficulty.

[0085] Smaller nanoparticles, with a pre-lyophilized size of approximately 100 nm or less, reduce tumors at lower dose concentrations compared to larger particles. For example, smaller nanoparticles achieve the same cancer therapeutic efficacy with only 1 / 5 the amount of taxane compared to larger nanoparticles. Therefore, lower doses of the drug can be used, and the risk of side effects is minimized.

[0086] Taxane-induced aggregates can also be linked to the target moiety or group to form conjugates, allowing the target group to bind to the nanocomposite particles of interest. Figures 10A-10B The targeting portion or group includes, but is not limited to, antibodies or fragments thereof (or their antigen-binding portions), antigens, homologous carbohydrates (e.g., sialic acid), cell surface receptor ligands, portions that bind to cell surface receptors (e.g., prostate-specific membrane antigen (PSMA)), portions that bind to cell surface sugars, extracellular matrix ligands, cytoplasmic receptor ligands, growth factors, cytokines, incretins, hormones, lectins, lectin targets such as galactose, galactose derivatives, N-acetylgalactosamine, mannose, mannose derivatives, vitamins such as folic acid, biotin, avidin, streptavidin, neutral avidin, DNA, RNA, etc.

[0087] Pharmaceutical formulation and nanoparticle preparation Taxane and the modified homopolymer can be separately suspended in suitable buffers and / or solvents (e.g., buffer solutions, methanol, acetone, ethanol, etc.) at appropriate concentrations (e.g., a defined concentration for in vivo use, typically in milligrams or nanograms). The two solutions are then mixed at a suitable temperature, such as room temperature, or at another temperature known to be acceptable for maintaining the integrity of the taxane and homopolymer, for a suitable period of time, such as 1 hour, 2 hours, etc. Other incubation times can range from a few minutes to several hours, as the aggregates of interest are stable once formed. The aggregates can be concentrated or collected by methods known in the art, such as by filtration, centrifugation, evaporation, lyophilization, dialysis, etc. The aggregates can be dried to extend shelf life.

[0088] For example, taxanes (e.g., paclitaxel) can be dissolved in methanol or ethanol in various amounts up to 40 mg / mL. (Hydrocarbon (CH3)(CH)2) 17 Modified random branched PEOX60 (monomer to initiator molar ratio = 60:1) was prepared as taught herein and dissolved in methanol or ethanol at different concentrations up to 100 mg / mL.

[0089] The two solutions were then mixed in different volumes to achieve a final homopolymer-to-taxane weight ratio in the range of 2:1 to 10:1, and rotary evaporated to dryness. The mixture was then redissolved in water or brine, aseptically filtered through a 0.22 μM filter, and freeze-dried by volume for 20 to 72 hours to obtain a dry powder.

[0090] Prior to lyophilization, the size range of the aggregates or nanoparticles, as measured by light scattering, can be approximately 50 nm to approximately 100 nm, approximately 60 nm to approximately 95 nm, and approximately 70 nm to approximately 90 nm (e.g., 3 mg paclitaxel / mL). After lyophilization, the size of the aggregates in diameter can range from approximately 110 nm to approximately 150 nm, approximately 115 nm to approximately 145 nm, and approximately 120 nm to approximately 140 nm (e.g., 5 mg paclitaxel / mL).

[0091] In one aspect, this disclosure relates to an aggregate comprising: a) a polyoxazoline comprising at least one first end group modified by a hydrophobic moiety, wherein the polyoxazoline further comprises a linear moiety, a branched moiety, or both, and the branched moiety comprises a symmetrically branched polymer, an asymmetricly branched polymer, or a combination thereof; and the polyoxazoline comprises a monomer to initiator molar ratio in the range of 50:1 to 80:1, and b) Taxane, The polyoxazoline and taxane comprise a polyoxazoline to taxane weight ratio of 6:1 to 8:1, and the aggregate size is from about 50 nm to about 100 nm. The aggregates include a filtration rate of 50% to 100% through a 0.22 µm filter.

[0092] Any of the above-described polyoxazoline polymers may be suitable. Aggregates may be formed as described herein.

[0093] The filtration efficiency of polymer-drug aggregates through a filter with a known pore size can be measured according to the following procedure: Polymer-drug aggregate samples can be dissolved in water, saline, PBS, or solvents as described herein at a predetermined final concentration.

[0094] Based on the filter composition, the polymer-aggregate can be dissolved in water, saline, PBS, other aqueous solutions, solvents, etc. In one example, a polymer-drug aggregate sample is prepared based on the weight of the polymer. In another example, a polymer-drug aggregate sample is prepared based on the weight of a drug, such as paclitaxel. In yet another example, a polymer-drug aggregate sample is prepared based on the final paclitaxel concentration (e.g., in mg / mL).

[0095] Samples can be passed through a filter by, for example, gravity or pressure. Pressure is typically applied to force the sample through the filter. An optional pressure gauge can be used. In one example, the container is sealed, and the sample is exposed to pressurized air. In another embodiment, the sealed container is configured to receive a vacuum in its collection section to extract the sample through an integral filter.

[0096] In this example, a syringe with a plunger is used in conjunction with a filter assembly that abuts against the nozzle portion of the syringe. A sample is loaded into the syringe barrel, and pressure is applied to the plunger to force the sample through the filter assembly. A constant pressure is applied to the plunger until all sample is expelled from the barrel. If filter clogging occurs, as evidenced by reduced back pressure and fluid flow through the filter into the collection section or collection container of the device, pressure is removed because excessive pressure on the plunger could rupture the membrane or potentially damage the assembly.

[0097] Load the predetermined sample volume V0 into the container. Then, attach the container to or expose it to the filter assembly. For a 0.22 μm filter assembly, a volume of 9–10 ml V0 can be selected. Record any remaining sample volume V1 in the container. Filter volume V f It can be used as formula V f =V0﹣V1 is calculated. Optionally, V f It can be determined by measuring the volume of the filtrate or the collected filtered sample.

[0098] Filtration rate R of the sample f The following formula can be used to calculate: R f =V f / V0.

[0099] R f It can be represented as a fraction, or a fraction can be converted to a percentage by multiplying it by 100.

[0100] It has been found that, for example, C 18 The polymer-drug aggregates prepared from PEOXABP60 and paclitaxel unexpectedly passed through a 0.22 μm filter with a filtration efficiency of 50%–100%. For comparison, the polymer-drug aggregates prepared from C… 18 The polymer-drug aggregates prepared from PEOXABP100 and paclitaxel have a filtration efficiency of less than 50%.

[0101] In the implementation plan, the filtration rate must be at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or greater; and must be within the range of 50%–100%, 60%–100%, 70%–100%, 80%–100%, 90%–100%, 55%–100%, 65%–100%, 75%–100%, 85%–100%, 95%–100%, 55%–95%, 6 The percentages must fall within the range of 5%–95%, 75%–95%, 85%–95%, 50%–95%, 60%–95%, 70%–95%, 80%–95%, 90%–95%, 50%–90%, 60%–90%, 70%–90%, 80%–90%, 55%–90%, 65%–90%, 75%–90%, and 85%–90%; or, in any case, must be 50% or greater.

[0102] In the implementation scheme, since the polymer-drug aggregates are intended for pharmaceutical use, and sterilization by filtration through a 0.22 μm filter is a common processing requirement or step, this finding of the correlation between the monomer:initiator ratio and the polymer:drug ratio, as well as the aggregate size, provides at least the advantage of ease of manufacture and reduced waste.

[0103] Choosing a specific monomer-to-initiator molar ratio and a specific polymer-to-drug ratio to produce aggregates of certain sizes is unexpected and surprising. Polyoxazoline polymers with lower monomer-to-initiator molar ratios, such as C... 18 -PEOXABP20 is unsuitable for pharmaceutical formulations due to manufacturing limitations and variations in cytotoxicity in some cell bioassays. Polymers with a monomer-to-initiator molar ratio of approximately 50:1 to 80:1 provide drug aggregates that offer advantages such as ease of manufacture, reduced cytotoxicity, and high filtration efficiency using 0.22 μm filters.

[0104] The first end group modified with the hydrophobic portion of the polyoxazoline polymer may comprise: a hydrophobic electrophilic molecule, including a hydrocarbon, aliphatic hydrocarbon, aromatic hydrocarbon, or a combination thereof; and a halide functional group, such as an alkyl halide, an aralkyl halide, an acyl halide, or a combination thereof; and may be provided by an initiator. Polymerization of the selected initiator and one or more monomers may produce a polyoxazoline of the present disclosure having at least one first end group modified with the hydrophobic portion. As disclosed throughout this application, the initiator may comprise a hydrophobic electrophilic molecule including a hydrocarbon. The hydrocarbon may comprise one to about 22 carbons, and the hydrocarbon may be saturated or unsaturated. In one embodiment, the initiator may comprise an aliphatic hydrocarbon, an aromatic hydrocarbon, or a combination thereof. The initiator may comprise a halide functional group. In one example, the initiator may comprise an alkyl halide, an aralkyl halide, an acyl halide, or a combination thereof.

[0105] Initiators may include alkyl halides containing one to about 22 carbons, including but not limited to methyl iodine, methyl bromide, methyl chloride, ethyl iodine, ethyl bromide, ethyl chloride, 1-iodopropane, 1-bromopropane, 1-chloropropane, 1-iodobutane, 1-bromobutane, 1-chlorobutane, 1-iodopentane, 1-bromopentane, 1-chloropentane, 1-iodohexane, 1-bromohexane, 1-chlorohexane, 1-iododecane, 1-bromododecane, 1-chlorododecane, 1-iodooctadecane, 1-bromooctadecane, 1-chlorooctadecane, and benzyl iodine, benzyl bromide, benzyl chloride, allyl bromide, acyl iodine, acyl bromide, acyl chloride, benzoyl bromide, or benzoyl chloride. In another example, the initiator comprises a p-toluenesulfonyl group.

[0106] As disclosed herein, the phrase "polyoxazoline includes the range of monomer to initiator molar ratios within which" means that a polyoxazoline is produced by reacting an initiator and at least one monomer in that specific range of monomer to initiator ratios (molar ratios) including both a start and an end point. Monomers such as oxazolines and substituted oxazolines disclosed herein may be suitable. Initiators disclosed above and below may be suitable.

[0107] Prior to lyophilization, the aggregates can range in size from 50 nm to approximately 100 nm. The term "size" refers to the particle size of the polymer-drug aggregate in solution, as measured using the dynamic light scattering method described below. The size of the polymer-drug aggregate can be measured in aqueous solution in one instance, in water in another instance, in saline solution in yet another instance, in a buffer solution such as phosphate-buffered saline (PBS) in yet another instance, and in a combination of saline and buffer solution in yet another instance.

[0108] In the aggregates disclosed herein, the polyoxazoline may further comprise a second end group containing a functional group modified with ethylenediamine (EDA) or an ethylenediamine derivative. The ethylenediamine derivative may include diethylenetriamine, triethylenetetramine, tetraethylenepentamine, pentaethylenehexamine, polyvinylamine, or tetramethylethylenediamine.

[0109] In the aggregates disclosed herein, taxanes can associate with at least one first end group modified with a hydrophobic portion. Taxanes can associate with the first end group via covalent, non-covalent, or a combination thereof. In one example, taxanes associate with a hydrocarbon via a non-covalent link. Not wishing to be bound by any particular theory or mechanism, the applicant believes that the hydrophobicity of the hydrocarbon in the polyoxazoline provides the desired interaction between the polymer and the taxane, which is water-insoluble or poorly soluble.

[0110] The aggregates disclosed herein may also include a targeting portion. The targeting portion may include an antibody or a fragment thereof, its antigen-binding portion, an antigen, a cell surface receptor, a cytoplasmic receptor, a cell receptor ligand, or a lectin ligand.

[0111] The polyoxazoline of this disclosure may include poly(2-oxazoline), poly(2-substituted oxazoline), or combinations thereof. In one example, the polyoxazoline may include poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-butyloxazoline), or combinations thereof. The polyoxazoline of this disclosure may be polymerized from at least one monomer including oxazoline, 2-substituted oxazoline, or combinations thereof. The oxazoline may be 2-oxazoline. The 2-substituted oxazoline may include 2-methyloxazoline, 2-ethyloxazoline, 2-propyloxazoline, 2-butyloxazoline, or combinations thereof.

[0112] In the aggregates of this disclosure, taxanes may include paclitaxel, docetaxel, or combinations thereof.

[0113] The aggregates disclosed herein may be sterile. In one instance, the aggregates may be sterilized by filtration through a 0.22 μm filter.

[0114] This disclosure also relates to a pharmaceutical composition comprising the aggregates disclosed herein. In some embodiments, the pharmaceutical composition comprises an aggregate comprising: a) a polyoxazoline comprising at least one first end group modified by a hydrophobic moiety, wherein the polyoxazoline further comprises a linear moiety, a branched moiety, or both, and the branched moiety comprises a symmetrically branched polymer, an asymmetricly branched polymer, or a combination thereof; and the polyoxazoline comprises a monomer to initiator molar ratio in the range of, for example, 50:1 to 80:1. b) Taxane, The polyoxazoline and taxane have a polyoxazoline to taxane weight ratio of, for example, 6:1 to 8:1, and the aggregate size is from about 50 nm to about 100 nm. The aggregates have a filtration efficiency of 50% to 100% through a 0.22 µm filter.

[0115] Polyoxazolines, as known or described herein, can be suitable polymers. Initiators for polyoxazolines can include hydrophobic, electrophilic molecules, such as hydrocarbons, including aliphatic hydrocarbons, aromatic hydrocarbons, or combinations thereof. The hydrocarbon can contain one to about 22 carbons and can be saturated or unsaturated. Initiators can contain halide functional groups, alkyl halides, aralkyl halides, acyl halides, or combinations thereof. In one example, the initiator may include methyl iodine, methyl bromide, methyl chloride, ethyl iodine, ethyl bromide, ethyl chloride, 1-iodopropane, 1-bromopropane, 1-chloropropane, 1-iodobutane, 1-bromobutane, 1-chlorobutane, 1-iodopentane, 1-bromopentane, 1-chloropentane, 1-iodohexane, 1-bromohexane, 1-chlorohexane, 1-iododecane, 1-bromododecane, 1-chlorododecane, 1-iodooctadecane, 1-bromooctadecane, 1-chlorooctadecane, benzyl iodine, benzyl bromide, benzyl chloride, allyl bromide, acyl iodine, acyl bromide, acyl chloride, benzoyl bromide, benzoyl chloride, or combinations thereof. In another example, the initiator comprises a p-toluenesulfonyl group. Polyoxazoline may also comprise a second end group containing a functional group modified with EDA or an ethylenediamine derivative, wherein the ethylenediamine derivative includes diethylenetriamine, triethylenetetramine, tetraethylenepentamine, pentaethylenehexamine, polyethyleneamine, or tetramethylethylenediamine. Polyoxazoline may include poly(2-oxazoline), poly(2-substituted oxazoline), or combinations thereof. In specific examples, polyoxazoline may include poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-butyloxazoline), or combinations thereof.

[0116] In a pharmaceutical composition, aggregates may include particles with a size of 50 nm to about 100 nm before lyophilization.

[0117] In the pharmaceutical composition, taxane can associate with at least one of the first end groups.

[0118] The aggregate may also contain a targeting portion, which, as disclosed above, may contain an antibody or a fragment thereof, its antigen-binding portion, an antigen, a cell surface receptor, a cytoplasmic receptor, a cell receptor ligand, or a lectin ligand.

[0119] In a pharmaceutical composition, taxane may include paclitaxel, docetaxel, or a combination thereof.

[0120] The pharmaceutical composition may also contain a carrier. The carriers disclosed herein may be suitable.

[0121] The pharmaceutical composition may be a cancer treatment for breast cancer, ovarian cancer, lung cancer, NSCLC (non-small cell lung cancer), colon cancer, gastric cancer, melanoma, head and neck cancer, pancreatic cancer, or combinations thereof. The pharmaceutical composition may be administered to the patient via parenteral, such as intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), mucosal, or rectal administration, or combinations thereof.

[0122] Unexpectedly, it was discovered that polyoxazoline polymers polymerized at specific ranges of monomer-to-initiator molar ratios offer filtration advantages for the manufacture of polymers having monomer-to-initiator molar ratios in the range of 50:1 to approximately 80:1, such as C 18 -PEOXABP60 can produce polymer-drug aggregates, which can achieve approximately 50% to 100% R... f A 0.22 μm membrane filter is used to produce sterile drug aggregates, while having, for example, a 100:1 (C 18 Polymers with monomers (-PEOXABP100) or larger in the molar ratio of monomer to initiator produce polymer-drug aggregates with a filtration efficiency of less than 50%. Such a low filtration efficiency leads to lower product yields, material waste, and longer filtration processes, resulting in reduced manufacturing productivity.

[0123] Without being bound by any particular theory or mechanism, polymer-drug aggregates produced from polymers with a monomer-initiator molar ratio of 100:1 or greater are believed to have undesirable aggregate sizes and distributions, and are susceptible to interparticle interactions, particle-filter interactions, or combinations thereof. Therefore, when particles are forced close to or near filter materials, such as when passing through a 0.22 μm filter under pressure, the aggregates may further aggregate or interact, leading to filter clogging and consequently lower filtration efficiency. Unexpectedly, monomer-initiator molar ratios, for example, in the range of 50:1 to 80:1, have been found to overcome the filter clogging problem, resulting in easier manufacturing, reduced waste, and increased productivity.

[0124] Pharmaceutical compositions of this disclosure for use as disclosed herein may be formulated to be compatible with the intended route of administration. Examples of routes of administration may include parenteral, intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), mucosal, or rectal administration. Solutions or suspensions for parenteral, intradermal, or subcutaneous application may include sterile diluents such as water for injection, saline, oil, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as EDTA; buffers such as acetate, citrate, or phosphate; and agents for adjusting tension, such as sodium chloride or dextran. The pH may be adjusted with an acid or base such as HCl or NaOH. Parenteral products may be packaged as articles in ampoules, disposable syringes, or multi-dose vials made of glass or plastic.

[0125] Suitable pharmaceutical compositions for injectable applications include sterile aqueous solutions or dispersions and sterile powders for immediate preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, antibacterial water, or phosphate-buffered saline (PBS). The composition is generally sterile and fluid to the extent necessary for injectability. The composition must be stable under manufacturing and storage conditions and must be protected against contamination by microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium comprising, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, liquid PEG, polysorbate, etc.), and suitable mixtures thereof. Appropriate flowability can be maintained, for example, by using coatings (e.g., lecithin), by maintaining the desired particle size in the case of dispersions, by using thickeners, and by using surfactants. Antimicrobial activity can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, etc. The composition may contain isotonic agents, such as sugars, polyols (e.g., mannitol, sorbitol), or sodium chloride. Prolonged absorption of injectable compositions can be caused by including agents that delay absorption, such as aluminum monostearate or gelatin, in the composition.

[0126] Sterile injectable solutions can be prepared by incorporating an active compound with one or a combination of the ingredients listed above into a suitable amount of solvent as needed, followed by filtration and sterilization. Typically, dispersions are prepared by incorporating the active compound into a sterile medium containing a base dispersion medium and other desired ingredients, such as those listed above, and as known in the art. In the case of sterile powders used to prepare sterile injectable solutions, the articles can be treated, for example, by lyophilization, vacuum drying, or freeze-drying, which produces a powder of the active ingredient plus any other desired components from its previously sterile filtered solution. Articles of interest can be stored and reconstituted with a suitable liquid for use.

[0127] Oral compositions typically contain an inert diluent, flavoring agent, odorant, or edible carrier. The composition may be encapsulated in gelatin capsules or compressed into tablets. For oral therapeutic administration, the active compound may be combined with excipients and administered in tablet, lozenge, or capsule form. Oral compositions may also be prepared using fluid carriers to produce syrups or liquid formulations, or used as mouthwashes, wherein the compound in the fluid carrier is administered orally and rinsed and spat out or swallowed.

[0128] Pharmaceutically compatible binders and / or excipients may be included as part of the composition. Tablets, pills, capsules, lozenges, etc., may contain binders such as microcrystalline cellulose, tragacanth gum, or gelatin; excipients such as starch or lactose; and disintegrants such as alginate or PRIMOGEL. ® (Modified corn starch, DFE pharma, DE trademark) or corn starch; lubricants, such as magnesium stearate or sterotes; gliding agents, such as colloidal silica; sweeteners, such as sucrose or saccharin; or flavorings, such as peppermint, methyl salicylate, or orange flavoring.

[0129] For administration by inhalation, the compound is delivered, for example, in the form of a wet or dry aerosol spray from a pressurized container or dispenser containing a suitable propellant, such as a gas, like carbon dioxide, or a spray or mist.

[0130] Systemic application can also be performed via mucosal or transdermal routes. For mucosal or transdermal application, a penetrant suitable for the barrier to be penetrated is used in the formulation. Such penetrants are generally known in the art and include, for example, detergents, bile salts, and fusidic acid derivatives for mucosal application. Mucosal application can be achieved by using nasal sprays or suppositories. For transdermal application, as is generally known in the art, the active compound is formulated as an ointment, cream, gel, or lotion. Suitable carriers include dimethyl sulfoxide.

[0131] The compound can also be prepared in the form of suppositories (e.g., using conventional suppository bases, such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

[0132] In one embodiment, the active compound is prepared using a carrier that will protect the compound from rapid elimination from the body, such as a controlled-release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used.

[0133] The methods for preparing such formulations will be apparent to those skilled in the art. Materials may also be commercially available, for example, from Alza Corporation and Nova Pharmaceuticals, Inc.

[0134] The aggregates of the present invention can be used topically, such as in the form of creams, ointments, lotions, oils, and other cosmetics. They may carry pharmaceutically active agents (PAAs), such as taxanes of interest and other bioactive or inert compounds, and the PAAs include emollients, bleaching agents, antiperspirants, pharmaceuticals, moisturizers, fragrances, colorants, pigments, dyes, antioxidants, oils, fatty acids, lipids, inorganic salts, organic molecules, sunscreens, vitamins, pharmaceuticals, keratolytic agents, UV blockers, tanning agents, bleaching agents, deodorants, fragrances, insect repellents, etc.

[0135] For ease of administration and consistency of dosage, it may be advantageous to formulate oral or parenteral compositions in unit dosage form. As used herein, “unit dosage form” means a physically discrete unit suitable as a unit dose for a subject to be treated; each unit contains a predetermined amount of the active compound calculated to produce the desired therapeutic endpoint.

[0136] Dosage, for example, preferred routes of administration and amounts, can be obtained based on empirical data from preclinical and clinical studies, and from practices known in the art. Dosage and delivery methods can be determined by and depend on the characteristics of the PAA, the polymer, the specific therapeutic effect to be achieved, the characteristics and condition of the recipient, etc. For repeated administration over several days or longer, depending on the condition, treatment can continue until the desired endpoint is reached.

[0137] This disclosure relates to a method for treating cancer in a subject with a corresponding need, the method comprising administering an effective dose of a first pharmaceutical composition to the subject; wherein the first pharmaceutical composition comprises an aggregate, the aggregate comprising: a) a polyoxazoline comprising at least one first end group modified by a hydrophobic moiety, wherein the polyoxazoline further comprises a linear moiety, a branched moiety, or both, the branched moiety comprising symmetrically branched polymers, asymmetrically branched polymers, or combinations thereof; and the polyoxazoline comprises a monomer to initiator molar ratio in the range of 50:1 to 80:1, and b) Taxane, and The subjects described herein have recurrent or metastatic cancer and have been treated with radiation therapy, surgery, chemotherapy, hormone therapy, biotherapy, immunotherapy, checkpoint inhibitor therapy, programmed death-ligand 1 (PD-L1)-based immune checkpoint inhibitor therapy, or a combination thereof, for at least 7 days prior to the first administration of the first drug composition.

[0138] In some cases, the subject may have recurrent or metastatic cancer and have been treated with radiation therapy, surgery, chemotherapy, hormone therapy, biotherapy, immunotherapy, checkpoint inhibitor therapy, programmed death-ligand 1 (PD-L1)-based immune checkpoint inhibitor therapy, or a combination thereof, for at least 14, 28, 35, 2, 3, or 6 months prior to the first administration of the first drug composition.

[0139] In some cases, the methods disclosed herein may also include the step of administering an effective dose of a second pharmaceutical composition to a subject simultaneously or sequentially with the first pharmaceutical composition, the second pharmaceutical composition comprising carboplatin, cisplatin, a monoclonal antibody or fragment thereof that binds to EGFR, a bispecific antibody or fragment thereof having at least one binding site for binding to EGFR, a trivalent antibody or fragment thereof having at least one binding site for binding to EGFR, a small molecule drug that binds to EGFR, or a combination thereof.

[0140] In some cases, the first and second pharmaceutical compositions may be formulated as single-dose units comprising the first and second pharmaceutical compositions, the single-dose units being selected from injectable packages of the first and second pharmaceutical compositions individually contained in a common package, premixed injectable packages comprising a mixture of the first and second pharmaceutical compositions, or combinations thereof.

[0141] In some cases, the second drug composition may be administered simultaneously with the first drug composition. It can be administered as a single injection using the combined first and second drug compositions. It can be administered as a combination of the first and second drug compositions via two separate injections. In some cases, the second drug composition containing cetuximab can be administered sequentially with the first drug composition. In some cases, the second drug composition may be administered to the subject within a range of 0.01 minutes to 7 days prior to administration of the first drug composition. In some cases, the second drug composition may be administered to the subject within a range of 0.01 minutes to 7 days after administration of the first drug composition.

[0142] In some cases, a single-dose unit may be an injectable package containing a first and a second pharmaceutical composition separately in a common package, wherein the first and second pharmaceutical compositions are packaged for injection into a subject over a predetermined time period ranging from 0.01 minutes to 10 hours or 7 days. In some cases, the first pharmaceutical composition is injected into the subject over a time period ranging from 0.01 minutes to 10 hours prior to the injection of the second pharmaceutical composition into the same subject. In some cases, the first pharmaceutical composition is injected into the subject over a time period ranging from 0.1 minutes to 10 hours after the injection of the second pharmaceutical composition into the same subject. This time period may range from 0.1 minutes to 10 hours, 0.1 minutes to 8 hours, 0.1 minutes to 6 hours, 0.1 minutes to 4 hours, 0.1 minutes to 3 hours, 0.1 minutes to 2 hours, 0.1 minutes to 1.5 hours, 0.1 minutes to 60 minutes, 0.1 minutes to 45 minutes, 0.1 minutes to 30 minutes, 0.1 minutes to 20 minutes, 0.1 minutes to 10 minutes, or 0.1 minutes to 5 minutes. In some cases, the injection package contains a first pharmaceutical composition and a second pharmaceutical composition in separate containers (e.g., vials, bottles, or tubes). In some cases, each container may be labeled for use as a first injection and a second injection. In some cases, the injection package may include instructions for use for injecting the first and second pharmaceutical compositions. In some cases, the injection package may include a sequential release device, wherein the first and second pharmaceutical compositions may be released in a defined order. In some cases, the second pharmaceutical composition may be released after the first pharmaceutical composition. In some cases, the second pharmaceutical composition may be released before the first pharmaceutical composition.

[0143] In some cases, a single-dose unit may be a premixed injectable package containing a mixture of a first pharmaceutical composition and a second pharmaceutical composition. The premixed injectable package may contain an effective dose of the first pharmaceutical composition and an effective dose of the second pharmaceutical composition mixed to form a single injectable unit.

[0144] In some cases, the second pharmaceutical composition may comprise carboplatin, cisplatin, a monoclonal antibody or fragment thereof that binds to EGFR, a bispecific antibody or fragment thereof having at least one binding site for EGFR, a trivalent antibody or fragment thereof having at least one binding site for EGFR, a small molecule drug that binds to EGFR, or a combination thereof. In some cases, the second pharmaceutical composition may comprise a trivalent antibody or fragment thereof having at least one binding site for EGFR. In some cases, the trivalent antibody or fragment thereof may comprise two bispecific binding sites against EGFR and one binding site against an additional target such as CD3. In some cases, the trivalent antibody or fragment thereof may comprise one binding site against EGFR and one or more binding sites against one or more additional targets.

[0145] In some cases, the monoclonal antibody or fragment thereof that binds to EGFR can be cetuximab.

[0146] In some cases, the effective dose of the first pharmaceutical composition may be contained in 15 mg / m². 2 Up to 500 mg / m 2 The range of taxanes, such as paclitaxel, and the effective dose of the second pharmaceutical composition may contain 100 mg / m². 2 Cisplatin.

[0147] In some cases, the effective dose of the first pharmaceutical composition may be contained in 15 mg / m². 2 Up to 500 mg / m 2 The range of taxanes, such as paclitaxel, and the effective dose of the second pharmaceutical composition may be contained in 50 mg / m². 2 Up to 900mg / m 2 Cetuximab within the range.

[0148] In some cases, any single-dose unit disclosed above and below may contain 15 mg / m³. 2 Up to 500 mg / m 2 A first pharmaceutical composition containing a taxane, such as paclitaxel, within the effective dose range, and may contain 100 mg / m³. 2 A second pharmaceutical composition containing an effective dose of cisplatin or carboplatin.

[0149] In some cases, any single-dose unit disclosed above and below may contain 15 mg / m³. 2 Up to 500 mg / m 2 A first pharmaceutical composition containing a taxane, such as paclitaxel, within the effective dose range, and may contain at 50 mg / m³. 2 Up to 900 mg / m2 A second pharmaceutical composition containing an effective dose of cetuximab within the range.

[0150] In some cases, any single-dose unit disclosed above and below may comprise a first pharmaceutical composition containing an effective dose of a taxane, such as paclitaxel, in the range of 30 mg to 1000 mg, and a second pharmaceutical composition containing an effective dose of cisplatin, such as 200 mg. In some cases, any single-dose unit disclosed above and below may comprise a first pharmaceutical composition containing an effective dose of a taxane, such as paclitaxel, in the range of 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, or 1000 mg, and a second pharmaceutical composition containing an effective dose of cisplatin, such as 200 mg.

[0151] In some cases, any single-dose unit disclosed above and below may comprise a first pharmaceutical composition containing an effective dose of a taxane, such as paclitaxel, in the range of 30 mg to 1000 mg, and a second pharmaceutical composition containing an effective dose of cetuximab in the range of 100 mg to 1800 mg. In some cases, any single-dose unit disclosed above and below may comprise a first pharmaceutical composition containing an effective dose of a taxane, such as paclitaxel, in the range of 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, or 1000 mg, and a second pharmaceutical composition containing an effective dose of cetuximab in the range of 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 1000 mg, 1500 mg, or 1800 mg.

[0152] Based on 1.07 m of human patients 2 Up to 1.9 m 2 (Approximately 1.7 m on average) 2 The average body surface area, and the single-dose unit of this disclosure, are applicable to administering 15 mg / m² to a patient. 2 Up to 500 mg / m 2 An effective dose of a first pharmaceutical composition containing taxane and may be contained in 50 mg / m³ 2 Up to 900 mg / m 2 Cetuximab within the range or 100 mg / m 2 A second pharmaceutical composition containing an effective dose of cisplatin.

[0153] In some cases, an effective dose of the first drug composition and an effective dose of the second drug composition may be administered to the subject at least once on days 1, 8, 15, 28, or a combination thereof during a treatment cycle. The term "treatment cycle" or "cycle" refers to a period of treatment for a subject or patient followed by a rest period, i.e., no treatment. In some cases, the drug composition is administered to the subject (i.e., the patient) on days 1, 8, and 15 (the treatment period) of a 28-day cycle (days 16–28 are a rest period without administration of the drug composition). Such a 28-day cycle can be a treatment cycle. In some cases, the drug composition is administered to the subject (i.e., the patient) on days 1, 8, 15, and 28 (the treatment period) of a 28-day treatment cycle, followed by a rest period ranging from 1 day to 3 months. Such a 28-day treatment and rest period can be a treatment cycle. The rest period allows the subject an opportunity to recover and build new healthy cells.

[0154] In some cases, the method may further include administering to a subject an alkaline aqueous solution, an oral alkalinizing agent, or a combination thereof, wherein the alkaline aqueous solution contains sodium bicarbonate in a percentage range of 0.01% to 10% based on the total weight of the alkaline aqueous solution, and may have a pH value in the range of 7.1 to 10.0. The alkaline aqueous solution may have a pH value in the range of 7.1 to 10.0, 7.1 to 9.5, 7.1 to 9.0, or 7.1 to 8.5. In an example, the alkaline aqueous solution may have a pH value in the range of 7.6 to 10.0, 7.6 to 9.5, 7.6 to 9.0, or 7.6 to 8.5. The alkaline aqueous solution or oral alkalinizing agent may be administered to the subject before, during, after, or in combination thereof, delivery of the nanoaggregates. The alkaline aqueous solution may be administered to the subject via intravenous (IV), subcutaneous (SC), intramuscular (IM), or intradermal (ID) injection, or a combination thereof. In one embodiment, the alkaline aqueous solution is administered to the subject via intravenous (IV), subcutaneous (SC), intramuscular (IM), or intradermal (ID) injection. Oral alkaline agents may include sodium bicarbonate tablets, solutions, powders, or granules. Commercially available products such as Brioschi (available from Neobourne Pharma LP., Quebec, Canada), Neut, or Sellymin may be suitable. Oral alkaline agents may be administered to the subject orally.

[0155] In some cases, the methods of this disclosure may include the step of administering sodium bicarbonate to a subject before, during, after, or in combination with the administration of FID-007. In some cases, the methods of this disclosure may include the step of administering to a subject an alkaline aqueous solution comprising a saline diluent (0.9% sodium chloride injection, USP) and 0.15% sodium bicarbonate (USP).

[0156] In some cases, the pharmaceutical composition may also contain sodium bicarbonate. The pharmaceutical composition may contain sodium bicarbonate in the range of 0.01% to 10% by weight of the total pharmaceutical composition. A sodium bicarbonate solution may comprise 0.15% sodium bicarbonate and 0.9% sodium chloride solution in water. With or without sodium chloride, the sodium bicarbonate solution may contain about 0.01% to 0.19% in one example, 0.1% to 0.19% in another example, 0.1% to 0.18% in yet another example, 0.1% to 0.15% in yet another example, 0.12% to 0.19% in yet another example, 0.12% to 0.18% in yet another example, 0.12% to 0.16% in yet another example, 4.2% in yet another example, 5% in yet another example, 7.5% in even yet another example, and 8.4% in yet another example. All percentages are based on the total weight of the pharmaceutical composition. As used herein, the term "drug" can refer to one or more of the bioactive agents disclosed herein. In one instance, a drug can be a taxane, such as paclitaxel.

[0157] In some cases, the aggregates may contain a polyoxazoline to taxane weight ratio of 6:1 to 8:1; and the size may be 70 nm to 90 nm.

[0158] In some cases, polyoxazoline may contain a second end group containing a functional group modified with ethylenediamine (EDA) or an EDA derivative, wherein the ratio of the second end group to EDA is 1:10.

[0159] In some cases, ethylenediamine derivatives can be configured to include diethylenetriamine, triethylenetetramine, tetraethylenepentamine, pentaethylenehexamine, polyethyleneamine, or tetramethylethylenediamine.

[0160] In some cases, the subject may have already been treated with at least one of the following drugs: afatinib, GILOTRIF ® (Afatinib), ABRAXANE ® (Albumin-bound paclitaxel or NAB-paclitaxel), bleomycin sulfate, carboplatin, cisplatin, cetuximab, docetaxel, ERBITUX ® (Cetuximab), Hydrea (hydroxyurea), KEYTRUDA ® (Pembrolizumab), Pembrolizumab, Methotrexate Sodium, Nivolumab, OPDIVO ® (Nivolumab), cisplatin, paclitaxel, TAXOL ® (paclitaxel), TAXOTERE ®(Docetaxel), Trexall (methotrexate sodium), a combination of carboplatin and Taxol, a combination of TPF (docetaxel, cisplatin, and fluorouracil (FU)) or a combination thereof (under the respective registered trademarks). In some cases, the subject may have already been treated with: pembrolizumab as a single agent with a combination positive score (CPS) ≥1; a combination of pembrolizumab, platinum, and fluorouracil; cetuximab; docetaxel; nivolumab; afatinib; or a combination thereof. Such treatment may be first-line or second-line treatment prior to treatment with the pharmaceutical compositions disclosed herein.

[0161] In some cases, the subject may have already received first-line treatment with: a combination of pembrolizumab, platinum, and fluorouracil; and second-line treatment with cetuximab, docetaxel, nivolumab, afatinib, or a combination thereof. Such treatment may be first-line, second-line, or both, prior to treatment with the pharmaceutical compositions disclosed herein.

[0162] In some cases, cancer can be breast cancer, ovarian cancer, lung cancer, NSCLC (non-small cell lung cancer), colon cancer, stomach cancer, melanoma, head and neck cancer (HNC), pancreatic cancer, or a combination thereof.

[0163] In some cases, head and neck cancer (HNC) can be squamous cell carcinoma of the head and neck (SCCHN), unresectable HNC, squamous cell carcinoma of the head and neck (HNSCC), human papillomavirus (HPV) positive head and neck cancer, metastatic HNC, metastatic HNC with 3 or more metastatic sites, metastatic squamous cell carcinoma with occult primary origin, pharyngeal cancer, hypopharyngeal cancer, laryngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, oral cancer, sinus cancer, nasal cavity cancer, salivary gland cancer, thyroid cancer, recurrent HNC, refractory HNC, HNC resistant to PD-1 therapy, HNC resistant to PD-L1 therapy, HNC resistant to immunotherapy, HNC resistant to T-cell therapy, HNC resistant to CAR-T therapy, HNC resistant to NK-cell therapy, and combinations thereof.

[0164] In some cases, initiators can be configured to include hydrophobic electrophilic molecules.

[0165] In some cases, the initiator can be configured to include hydrocarbons.

[0166] In some cases, the initiator can be configured to include aliphatic hydrocarbons, aromatic hydrocarbons, or a combination of both.

[0167] In some cases, the initiator is configured to contain halide functional groups.

[0168] In some cases, the initiator can be configured to include alkyl halides, aralkyl halides, acyl halides, or combinations thereof.

[0169] In some cases, the hydrocarbon can be configured to contain 1 to 22 carbons, and the hydrocarbon can be saturated or unsaturated.

[0170] In some cases, the initiator may be configured to include methyl iodine, methyl bromide, methyl chloride, ethyl iodine, ethyl bromide, ethyl chloride, 1-iodopropane, 1-bromopropane, 1-chloropropane, 1-iodobutane, 1-bromobutane, 1-chlorobutane, 1-iopentane, 1-bromopentane, 1-chloropentane, 1-iohexane, 1-bromohexane, 1-chlorohexane, 1-iododecane, 1-bromododecane, 1-chlorododecane, 1-iooctadecane, 1-bromooctadecane, 1-chlorooctadecane, benzyl iodine, benzyl bromide, benzyl chloride, allyl bromide, acyl iodine, acyl bromide, acyl chloride, benzoyl bromide, benzoyl chloride, or combinations thereof.

[0171] In some cases, the initiator can be configured to contain a p-toluenesulfonyl group.

[0172] In some cases, taxanes can be configured to associate with at least one first end group.

[0173] In some cases, polyoxazoline can be configured to include poly(2-oxazoline), poly(2-substituted oxazoline), or combinations thereof.

[0174] In some cases, polyoxazoline can be configured to include poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), or poly(2-butyloxazoline) or combinations thereof.

[0175] In some cases, taxanes can be configured to include paclitaxel, docetaxel, or a combination thereof.

[0176] In some cases, the aggregates may contain a 7:1 ratio of polyoxazoline to taxane.

[0177] In some cases, the pharmaceutical composition may be administered to the subject via parenteral administration, wherein the parenteral administration is selected from intravenous (IV) injection, intradermal injection, subcutaneous injection, or a combination thereof.

[0178] In some cases, aggregates can be configured to also include a target portion.

[0179] In some cases, the targeting portion can be configured to include an antibody or a fragment thereof, its antigen-binding portion, an antigen, a cell surface receptor, a cytoplasmic receptor, a cell receptor ligand, or a lectin ligand.

[0180] In some cases, the methods of this disclosure may include steps for monitoring treatment progress in subjects. Treatment can be monitored using conventional techniques and assays, such as blood tests, CT scans, imaging, and patient input.

[0181] Suitable methods of this disclosure may include pharmaceutical compositions in containers, packages, or dispensers, such as one of the single-dose units disclosed herein, along with instructions for administration.

[0182] In some cases, this disclosure relates to a pharmaceutical composition comprising: a first pharmaceutical composition comprising: a) a polyoxazoline comprising at least one first end group modified by a hydrophobic moiety, wherein the polyoxazoline further comprises a linear moiety, a branched moiety, or both, the branched moiety comprising an asymmetrically branched polymer; and the polyoxazoline comprises a monomer to initiator molar ratio of 60:1. b) Taxane, The first pharmaceutical composition comprises a polyoxazoline to taxane weight ratio of 6:1 to 8:1; and has a size of 70 nm to 90 nm; and the polyoxazoline comprises a second end group containing a functional group modified with ethylenediamine (EDA) or an EDA derivative, wherein the ratio of the second end group to EDA is 1:10; and The second pharmaceutical composition comprises carboplatin, cisplatin, a monoclonal antibody or fragment thereof that binds to EGFR, a bispecific antibody or fragment thereof having at least one binding site for binding to EGFR, a trivalent antibody or fragment thereof having at least one binding site for binding to EGFR, a small molecule drug that binds to EGFR, or a combination thereof. The pharmaceutical composition is formulated as a single-dose unit, the single-dose unit comprising a first pharmaceutical composition and a second pharmaceutical composition containing taxane, the single-dose unit being selected from an injection package containing a first pharmaceutical composition and a second pharmaceutical composition separately in a common package for injection into a subject over a predetermined time period, a premixed injection package containing a mixture of the first pharmaceutical composition and the second pharmaceutical composition, or a combination thereof.

[0183] In some cases, monoclonal antibodies or fragments thereof that bind to EGFR may include cetuximab.

[0184] In some cases, the pharmaceutical composition may also contain a salt component containing sodium bicarbonate, and the pharmaceutical composition may have a pH value in the range of about 7.1 to about 10.

[0185] In some cases, a single-dose unit may be an injectable package containing a first and a second pharmaceutical composition separately in a common package, wherein the first and second pharmaceutical compositions are packaged for injection into a subject over a predetermined time period ranging from 0.01 minutes to 10 hours or 7 days. In some cases, the first pharmaceutical composition is injected into the subject over a time period ranging from 0.01 minutes to 10 hours prior to the injection of the second pharmaceutical composition into the same subject. In some cases, the first pharmaceutical composition is injected into the subject over a time period ranging from 0.1 minutes to 10 hours after the injection of the second pharmaceutical composition into the same subject. This time period may range from 0.1 minutes to 10 hours, 0.1 minutes to 8 hours, 0.1 minutes to 6 hours, 0.1 minutes to 4 hours, 0.1 minutes to 3 hours, 0.1 minutes to 2 hours, 0.1 minutes to 1.5 hours, 0.1 minutes to 60 minutes, 0.1 minutes to 45 minutes, 0.1 minutes to 30 minutes, 0.1 minutes to 20 minutes, 0.1 minutes to 10 minutes, or 0.1 minutes to 5 minutes. In some cases, the injection package contains a first pharmaceutical composition and a second pharmaceutical composition in separate containers (e.g., vials, bottles, or tubes). In some cases, each container may be labeled for use as a first injection and a second injection. In some cases, the injection package may include instructions for use for injecting the first and second pharmaceutical compositions. In some cases, the injection package may include a sequential release device, wherein the first and second pharmaceutical compositions may be released in a defined order. In some cases, the second pharmaceutical composition may be released after the first pharmaceutical composition. In some cases, the second pharmaceutical composition may be released before the first pharmaceutical composition.

[0186] In some cases, a single-dose unit may be a premixed injectable package containing a mixture of a first pharmaceutical composition and a second pharmaceutical composition. The premixed injectable package may contain an effective dose of the first pharmaceutical composition and an effective dose of the second pharmaceutical composition mixed to form a single injectable unit.

[0187] In some cases, the second pharmaceutical composition may comprise carboplatin, cisplatin, a monoclonal antibody or fragment thereof that binds to EGFR, a bispecific antibody or fragment thereof having at least one binding site for binding to EGFR, a trivalent antibody or fragment thereof having at least one binding site for binding to EGFR, a small molecule drug that binds to EGFR, or a combination thereof.

[0188] In some cases, the monoclonal antibody or fragment thereof that binds to EGFR can be cetuximab.

[0189] In some cases, the effective dose of the first pharmaceutical composition may be contained in 15 mg / m². 2 Up to 500 mg / m 2The range of taxanes, such as paclitaxel, and the effective dose of the second pharmaceutical composition may contain 100 mg / m². 2 Cisplatin.

[0190] In some cases, the effective dose of the first pharmaceutical composition may be contained in 15 mg / m². 2 Up to 500 mg / m 2 The range of taxanes, such as paclitaxel, and the effective dose of the second pharmaceutical composition may be contained in 50 mg / m². 2 Up to 900mg / m 2 Cetuximab within the range.

[0191] In some cases, any single-dose unit disclosed above and below may contain 15 mg / m³. 2 Up to 500 mg / m 2 A first pharmaceutical composition containing a taxane, such as paclitaxel, within the effective dose range, and may contain 100 mg / m³. 2 A second pharmaceutical composition containing an effective dose of cisplatin.

[0192] In some cases, any single-dose unit disclosed above and below may contain 15 mg / m³. 2 Up to 500 mg / m 2 A first pharmaceutical composition containing a taxane, such as paclitaxel, within the effective dose range, and may contain at 50 mg / m³. 2 Up to 900 mg / m 2 A second pharmaceutical composition containing an effective dose of cetuximab within the range.

[0193] In some cases, any single-dose unit disclosed above and below may comprise a first pharmaceutical composition containing an effective dose of a taxane, such as paclitaxel, in the range of 30 mg to 1000 mg, and a second pharmaceutical composition containing an effective dose of cisplatin, such as 200 mg. In some cases, any single-dose unit disclosed above and below may comprise a first pharmaceutical composition containing an effective dose of a taxane, such as paclitaxel, in the range of 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, or 1000 mg, and a second pharmaceutical composition containing an effective dose of cisplatin, such as 200 mg.

[0194] In some cases, any single-dose unit disclosed above and below may comprise a first pharmaceutical composition containing an effective dose of a taxane, such as paclitaxel, in the range of 30 mg to 1000 mg, and a second pharmaceutical composition containing an effective dose of cetuximab in the range of 100 mg to 1800 mg. In some cases, any single-dose unit disclosed above and below may comprise a first pharmaceutical composition containing an effective dose of a taxane, such as paclitaxel, in the range of 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, or 1000 mg, and a second pharmaceutical composition containing an effective dose of cetuximab in the range of 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 1000 mg, 1500 mg, or 1800 mg.

[0195] Based on 1.07 m of human patients 2 Up to 1.9 m 2 (Approximately 1.7 m on average) 2 The average body surface area, and the single-dose unit of this disclosure, are applicable to administering 15 mg / m² to a patient. 2 Up to 500 mg / m 2 An effective dose of a first pharmaceutical composition containing taxane and may be contained in 50 mg / m³ 2 Up to 900 mg / m 2 Cetuximab within the range or 100 mg / m 2 A second pharmaceutical composition containing an effective dose of cisplatin.

[0196] In some cases, this disclosure relates to the use of a first pharmaceutical composition and a second pharmaceutical composition in the manufacture of a medicament for treating head and neck cancer in a subject with a corresponding need, wherein, The first pharmaceutical composition comprises: a) a polyoxazoline comprising at least one first end group modified by a hydrophobic moiety, wherein the polyoxazoline further comprises a linear moiety, a branched moiety, or both, the branched moiety comprising an asymmetrically branched polymer; and the polyoxazoline comprises a monomer to initiator molar ratio of 60:1. b) Taxane, The first pharmaceutical composition comprises a polyoxazoline to taxane weight ratio of 6:1 to 8:1; and has a size of 70 nm to 90 nm; and the polyoxazoline comprises a second end group containing a functional group modified with ethylenediamine (EDA) or an EDA derivative, wherein the ratio of the second end group to EDA is 1:10; and The second pharmaceutical composition comprises carboplatin, cisplatin, a monoclonal antibody or fragment thereof that binds to EGFR, a bispecific antibody or fragment thereof having at least one binding site for binding to EGFR, a trivalent antibody or fragment thereof having at least one binding site for binding to EGFR, a small molecule drug that binds to EGFR, or a combination thereof. The drug is formulated as a single-dose unit comprising the first drug composition and the second drug composition, the single-dose unit being selected from injectable packages containing the first drug composition and the second drug composition separately in a common package, premixed injectable packages containing a mixture of the first drug composition and the second drug composition, or combinations thereof.

[0197] In some cases, monoclonal antibodies or fragments thereof that bind to EGFR may include cetuximab.

[0198] In some cases, as disclosed above and below, the subject is administered an effective dose of the first pharmaceutical composition and an effective dose of the second pharmaceutical composition at least once on day 1, day 8, day 15, day 28, or a combination thereof.

[0199] The single-dose units and administration (injection) methods and times disclosed herein are applicable to the uses described herein.

[0200] In some cases, this disclosure also relates to methods of administering one or more of the aggregates or pharmaceutical compositions disclosed herein as a food supplement or additive, or as a vitamin-like dosage form for preventative use, into or with a food or beverage. The aggregates of interest may be encapsulated in a form that will survive passage through the gastric environment. Such forms are well known, for example, enteric-coated formulations. Alternatively, as known in the art, the aggregates of interest may be modified to enhance their half-life, for example, through chemical modification or combination with agents known to cause delayed, sustained, or controlled release.

[0201] The present disclosure will now be illustrated by examples in the following non-limiting embodiments.

[0202] Example This disclosure is further defined in the following embodiments. It should be understood that while the embodiments indicate implementations of this disclosure, they are given by way of illustration only. Based on the foregoing discussion and these embodiments, those skilled in the art can determine the essential features of this disclosure, and various changes and modifications can be made to this disclosure to suit various uses and conditions without departing from the spirit and scope of this disclosure.

[0203] Material Symmetrically branched PPI dendrimers were purchased from Sigma-Aldrich. Symmetrically branched PEI dendrimers and dendritic grafts were prepared according to known procedures. All antibodies were purchased from Sigma-Aldrich, Biodesign, or Fitzgerald. Different generations of PAMAM dendrimers were purchased from Dendritech, Inc.

[0204] Example 1 Symmetrically branched PPI (m﹣SB﹣PPI﹣NH) modified with amino functional groups 2 ﹣1.0) The following reagents were used: symmetrically branched PPI (SB-PPI-4, 8, 16, 32, 64, MW 316, 773, 1,687, 3,514 and 7,168), methyl acrylate (MA, FW=86.09), EDA (FW=60.10) and methanol.

[0205] Add 1.0 g of PPI-64 dendritic polymer (MW 7168) and 20 ml of methanol (solution A) to a round-bottom flask. Add 2.4 g of methyl acrylate (MA) and 10 ml of methanol (solution B) to a separate round-bottom flask. Then, while stirring at room temperature, slowly add solution A dropwise to solution B. Allow the resulting solutions to react at 40°C for 2 hours. After the reaction is complete, remove the solvent and unreacted MA monomer by rotary evaporation, and then redissolve 2.5 g of MA-functionalized PPI in 20 ml of methanol.

[0206] 160 g of EDA and 50 ml of methanol were added to a round-bottom flask at 0 °C, followed by the slow addition of MA-functionalized PPI. The solution was then allowed to react at 4 °C for 48 hours. The solvent and excess EDA were removed by rotary evaporation. The crude product was then precipitated from the ether solution and further purified by dialysis to give approximately 2.8 g of primary amine-functionalized symmetrically branched PPI (m﹣SB﹣PPI﹣NH2﹣1.0) with a molecular weight of approximately 21,760. 1 H and 13 The products were characterized by nuclear magnetic resonance (NMR) and size exclusion chromatography (SEC).

[0207] Symmetrically branched PPI dendritic polymers and symmetrically branched PEI dendritic grafts with various molecular weights modified with other MA or primary amines were prepared in a similar manner.

[0208] Example 2 Symmetrically branched PPI (mix﹣m﹣SB﹣PPI﹣64﹣NH) modified with mixed hydroxyl and amino functional groups 2 / OH﹣2) The amino-functionalized symmetrically branched PPI (m-SB-PPI-64-NH2-1.0), MA, EDA, monoethanolamine (MEA, FW=61.08) and methanol were utilized.

[0209] Add 1.0 g of amino-modified PPI or m-SB-PPI-NH2-1.0 produced by the previous procedure and 20 ml of methanol (solution A) to a round-bottom flask. Add 2.4 g of MA and 10 ml of methanol to a separate round-bottom flask (solution B). Then, while stirring at room temperature, slowly add solution A dropwise to solution B. Allow the resulting solutions to react at 40°C for 2 hours. After the reaction is complete, remove the solvent and unreacted monomer MA by rotary evaporation, and then redissolve 2.5 g of the product, MA-functionalized m-SB-PPI-64-MA-1.5, in 20 ml of methanol.

[0210] 32 g EDA, 130 g MEA, and 100 ml methanol (EDA:MEA molar ratio of 20:80) were added to a round-bottom flask at 0 °C, followed by the slow addition of SB-PPI-64-MA-1.5. The solution was then allowed to react at 4 °C for 48 hours. The solvent and excess EDA were removed by rotary evaporation. The crude product was then precipitated from the ether solution and further purified by dialysis to give approximately 2.8 g of a mixed hydroxyl and amino-functionalized (mixed surface) SBP (mix-m-SB-PPI-64-NH2 / OH-2.0, with an average of 20% NH2 and 80% OH surface groups and a molecular weight of approximately 21,862).

[0211] Random AB-PEI and regular AB-PLL molecules with different ratios of hydroxyl and amino groups and different molecular weights were prepared in a similar manner.

[0212] Randomly branched PEIs were purchased from Aldrich and Polysciences. ABPs were prepared according to known procedural rules. All antibodies were purchased from Sigma-Aldrich, Biodesign, or Fitzgerald.

[0213] Example 3 Randomly branched PEI(m﹣ran﹣AB﹣PEI﹣NH) modified with amino functional groups 2 ﹣1.0) Using randomly asymmetric branched PEI (ran﹣AB﹣PEI, MW 2,000, 25,000 and 75,000), MA, EDA and methanol.

[0214] Add 1.0 g PEI (MW 2,000) and 20 mL methanol (solution A) to a round-bottom flask. Add 3.0 g MA and 10 mL methanol (solution B) to a separate round-bottom flask. Then, while stirring at room temperature, slowly add solution A dropwise to solution B. Allow the resulting solution to react at 40 °C for 2 hours. After the reaction is complete, remove the solvent and unreacted MA by rotary evaporation, and then redissolve the MA-functionalized PEI in 20 mL of methanol.

[0215] 80 g of EDA and 50 ml of methanol were added to a round-bottom flask at 0 °C, followed by the slow addition of MA-functionalized PEI (1 g of MA dissolved in 20 ml of methanol). The solution was then allowed to react at 4 °C for 48 hours. The solvent and excess EDA were removed by rotary evaporation. The crude product was then precipitated from the ether solution and further purified by dialysis to give approximately 3.0 g of primary amine-functionalized, randomly asymmetricly branched PEI (m﹣ran﹣AB﹣PEI﹣NH2﹣1.0) with a molecular weight of approximately 7,300. 1 H NMR and 13 The products were characterized by C NMR and SEC.

[0216] Randomly asymmetric branched PEI and regularly asymmetric branched PLL polymers with various molecular weights modified by other MA or primary amines were prepared in a similar manner.

[0217] Example 4 Modification of branched polymers with hydrocarbon chains Modification of randomly branched PEI with 10% hydrocarbon chain was used as an example. 1 gram of branched PEI (FW=25000) was dissolved in 10 mL of methanol. 0.23 g of 1,2-epoxyhexane (FW=100.16) was added to the solution, and the mixture was heated at 40 °C for 2 hours. The solvent was then rotary evaporated, and the residue was redissolved in water. After dialysis (cutoff value 3,500), modified PEI was obtained.

[0218] Prepare in a similar manner those having various percentages and lengths (e.g., C4, C...). 12 C 18 and C 22 Other MBPs of the hydrocarbon chain, such as PAMAM, PEI and PPI dendritic polymers and dendritic grafts, as well as asymmetric PLLs.

[0219] Example 5 Randomly asymmetric branched PEI (m﹣ran﹣AB﹣PEI﹣) modified with mixed hydroxyl and amino functional groups NH 2 / OH﹣2) The amino-functionalized random asymmetric branched PEI (m﹣ran﹣AB﹣PEI﹣NH2﹣1.0), MA, EDA, monoethanolamine (MEA, FW=61.08) and methanol were utilized.

[0220] Add 1.0 g of amino-modified PEI or m﹣ran﹣AB﹣PEI﹣NH2﹣1.0 produced by the previous procedure and 20 ml of methanol (solution A) to a round-bottom flask. Add 3.0 g of MA and 10 ml of methanol to a separate round-bottom flask (solution B). Then, while stirring at room temperature, slowly add solution A dropwise to solution B. Allow the resulting solutions to react at 40 °C for 2 hours. After the reaction is complete, remove the solvent and unreacted MA by rotary evaporation, and then redissolve the product MA-functionalized m﹣ran﹣AB﹣PEI﹣MA﹣1.5 in 20 ml of methanol.

[0221] 60 g EDA, 244 g MEA, and 100 ml methanol (EDA:MEA molar ratio of 20:80) were added to a round-bottom flask at 0 °C, followed by the slow addition of m﹣ran﹣AB﹣PEI﹣MA﹣1.5 (1 g MA dissolved in 20 ml methanol). The solution was then allowed to react at 4 °C for 48 hours. The solvent and excess EDA were removed by rotary evaporation. The crude product was then precipitated from the ether solution and further purified by dialysis to give approximately 2.4 g of a mixed hydroxyl and amino-functionalized random ABP (m﹣ran﹣AB﹣PEI﹣NH2 / OH﹣2.0, with an average of 20% NH2 and 80% OH surface groups and a molecular weight of approximately 18,000).

[0222] Random AB-PEI polymers and regular AB-PLL polymers with various ratios of hydroxyl and amino groups and other modifications of different molecular weights were prepared in a similar manner.

[0223] Example 6 Randomly branched poly(2-ethyloxazoline) (PEOX) with alkyl-modified terminal groups of primary amine chains. CH3﹣(CH2) 11 -PEOX-ABP100(C 12 ABP100 (an arbitrarily chosen name to indicate the molar ratio of monomer to initiator in the initial reaction) provides a general procedure for the preparation of core-shell structures. CH3(CH2) 11A mixture of -Br (2.52 g) in 500 mL of toluene was azeotropically heated under N2 to remove water using a distillation head for approximately 15 min. 2-Ethyloxazoline (100 g) was added dropwise through a feeding funnel, allowing the mixture to reflux between 24 and 48 hours. After polymerization was complete, 12.12 g of EDA was added to the reactive polymer solution (A) to introduce amine functional groups. The molar ratio of POX chain ends to EDA was 1:20.

[0224] Alternatively, N-Boc-piperazine or water (e.g., with 1 N Na₂CO₃) can be added to terminate the reaction. Morpholine or PEI can also be added to the reactive polymer solution (A) to terminate the reaction. The crude product is redissolved in methanol and then precipitated from a large excess of diethyl ether. The bottom layer is redissolved in methanol and dried by rotary evaporation and vacuum drying to give an asymmetricly randomized PEOX polymer as a white solid (101 g).

[0225] Other asymmetrically randomly branched polymers, such as C6-PEOX ABP20, 50, 100, 200, 300, and 500, were prepared in a similar manner. 12 -PEOX ABP20, 50, 200, 300 and 500, C 22 -PEOX ABP20, 50, 100, 200, 300 and 500, as well as polystyrene-PEOX, and unmodified and modified poly(2-substituted oxazoline), such as poly(2-methyloxazoline). All products were analyzed by SEC and NMR.

[0226] Example 7 Randomly branched poly(2-ethyloxazoline) (PEOX) with alkyl-modified terminal groups of primary amine chains. CH3﹣(CH2) 17 -PEOX-ABP60(C 18 -PEOXABP60 is an arbitrary name to indicate the molar ratio of monomer to initiator in the initial reaction. The synthesis of this material provides a general procedure for preparing core-shell structures. CH3(CH2) 17 A mixture of -Br (5.61 g) in 500 ml of toluene was azeotropically heated under N2 to remove water using a distillation head for approximately 15 min. 2-Ethyloxazoline (100 g) was added dropwise through a feeding funnel, allowing the mixture to reflux between 24 and 48 hours. After polymerization was complete, 10.1 g of EDA was added to the reactive polymer solution (A) to introduce amine functional groups. The molar ratio of the polyoxazoline reactive chain terminus to EDA was 1:10.

[0227] At a ratio of 1:10, the size of the nanoaggregates is reduced, making them easier to filter and resulting in less filter clogging during manufacturing. It also reduces the amount of hydrolysis byproducts and improves the purity of the final drug product.

[0228] Alternatively, N-Boc-piperazine or water (e.g., with 1 N Na₂CO₃) can be added to terminate the reaction. Morpholine or PEI can also be added to the reactive polymer solution (A) to terminate the reaction. The crude product is redissolved in methanol and then precipitated from a large excess of diethyl ether. The bottom layer is redissolved in methanol and dried by rotary evaporation and vacuum drying to give an asymmetric, randomly branched PEOX polymer as a white solid.

[0229] Other asymmetric, randomly branched polymers, such as C, can be prepared in a similar manner. 18 -PEOX ABP20, 40, 50, 70, 80, 100, 120, 200, 300, 500, etc., as well as unmodified and modified poly(2-substituted oxazoline), such as poly(2-methyloxazoline). All products were analyzed by SEC and NMR.

[0230] Example 8 Hybrid surface-modified symmetrically branched polymer-IgG conjugates The preparation of symmetrically branched PPI-IgG conjugates (mix-m-SB-PPI-64-NH2 / OH-2-IgG conjugates) modified with mixed surfaces (OH / NH2 mix) is provided as a general procedure for the preparation of polymer antibodies.

[0231] Other conjugates were also obtained in a similar manner, such as m﹣SB﹣PPI﹣4﹣NH2﹣1﹣IgG, m﹣SB﹣PPI﹣8﹣NH2﹣1﹣IgG, m﹣SB﹣PPI﹣16﹣NH2﹣1﹣IgG, m﹣SB﹣PPI﹣32﹣NH2﹣1﹣IgG, m﹣SB﹣PPI﹣4﹣NH2﹣2﹣IgG, m﹣SB﹣PPI﹣8﹣NH2﹣2﹣IgG, m﹣SB﹣PP I-16-NH2-2-IgG, m-SB-PPI-32-NH2-2-IgG, m-SB-PPI-4-NH2-3-IgG, m-SB-PPI-8-NH2-3-I gG, m﹣SB﹣PPI﹣16﹣NH2﹣3﹣IgG, m﹣SB﹣PPI﹣32﹣NH2﹣3﹣IgG, mix﹣m﹣SB﹣PPI﹣4﹣NH2 / OH﹣1(OH / NH2 mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣8﹣NH2 / OH﹣1(OH / NH2 mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣16﹣NH2 / OH﹣1(OH / NH2 mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣32﹣NH2 / OH﹣1(OH / NH2 mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣4﹣NH2 / OH﹣2(OH / NH2mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣8﹣NH2 / OH﹣2(OH / NH2 mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣16﹣NH2 / OH﹣2(OH / NH2 mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣32﹣NH2 / OH﹣2(OH / NH2mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣4﹣NH2 / OH﹣3(OH / NH2 mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣8﹣NH2 / OH﹣3(OH / NH2mix)﹣IgG, mix﹣m﹣SB﹣PPI﹣16﹣NH2 / OH﹣3(OH / NH2 Mix()﹣IgG, mix﹣m﹣SB﹣PPI﹣32﹣NH2 / OH﹣3(OH / NH2mix)﹣IgG, and comb-like burst PEI dendritic grafts modified with primary amines and mix OH / NH2 (generations 0-5). Other targeting moieties attached to the modified SBP of interest were also synthesized in a similar manner.

[0232] Example 9 LC﹣SPDP﹣Mixed Surface m﹣SB﹣PPI﹣64﹣NH 2 / OH﹣2 The randomized mixed surface components mix﹣m﹣SB﹣PPI﹣64﹣NH2 / OH﹣2 (4×10⁻⁶) were added to 400 μl of phosphate buffer (20 mM phosphate and 0.1 M NaCl, pH 7.5). ﹣7 4 × 10 mol) added to 400 μL of water ﹣6 mol sulfonyl-LC-SPDP (Pierce, IL). The mixture was vortexed and incubated at 30 °C for 30 min. LC-SPDP-mix-m-SB-PPI-64-NH2 / OH-2 was purified by gel filtration chromatography and equilibrated with buffer A (0.1 M phosphate, 0.1 M NaCl and 5 mM EDTA, pH 6.8). The product was further concentrated to give 465 μL of a solution with a concentration of approximately 0.77 nmol.

[0233] Example 10 Thiolized mix﹣m﹣SB﹣PPI﹣64﹣NH 2 / OH﹣2 LC-SPDP mix-m-SB-PPI-64-NH2 / OH-2 (50 nmol in 65 μl of buffer A) was mixed with 100 μL of dithiothreitol (DTT) (50 mM in buffer A) and incubated at room temperature for 15 min. Excess DTT and byproducts were removed by gel filtration through buffer A. The product was concentrated in a 10 K Centricon concentrator to obtain 390 μL of the thiolized mix-m-SB-PPI-64-NH2 / OH-2, which was used for conjugation with activated antibodies.

[0234] Example 11 Maleimide R (MAL-R)-activated antibody Add 20.4 μL of MAL﹣R﹣NHS (N-hydroxysuccinimide) solution (10 mM in water) to the antibody (310 μL, 5.1 mg, or 34 nmol) in PBS. Vortex the mixture and incubate at 30 °C for 15 min. Purify the product by gel filtration using buffer A. The maleimide﹣R﹣ activated antibody is used for conjugation with thiolized mix﹣m﹣SB﹣PPI﹣64﹣NH2 / OH﹣2.

[0235] Example 12 mix﹣m﹣SB﹣PPI﹣64﹣NH 2 / OH﹣2﹣antibody conjugate Add MAL-R activated antibody (4.8 mL, or 34 nmol) to the thiolized mix-m-SB-PPI-64-NH2 / OH-2 (310 μL or 35.7 nmol). Concentrate the reaction mixture to approximately 800 μL and then allow it to incubate overnight at 4 °C and / or at room temperature for approximately 1 hour. After completion, quench the reaction with 100 μL of ethylmaleimide (50 mM solution), and then fractionate the conjugate on a carboxymethyl cellulose (CM cellulose) column (5 mL) using a sodium chloride gradient in 20 mM phosphate buffer at pH 6. The conjugate was eluted with a sodium chloride gradient and characterized by cation exchange chromatography, UV spectroscopy, and polyacrylamide gel electrophoresis.

[0236] Example 13 via antibody reductive coupling – reductive conjugation Add 40 μL of DTT (50 mM in buffer B) to 160 μL of 2.1 mg or 14 nmol of antibody in buffer B (containing 0.1 M sodium phosphate, 5 mM EDTA, and 0.1 M NaCl, pH 6.0). Allow the solution to stand at room temperature for 30 min. Purify the product by gel filtration in a Sephadex G-25 column equilibrated with buffer B. Concentrate the reduced antibody to 220 μL and use it for conjugation.

[0237] Example 14 MAL-R-SBP with mixed surface modification Add 400 μL (400 × 10⁻⁶) to pH 7.4. ﹣9 400 μL of MAL-R-NHS (10 mM in water) was added to a mixture of surface-modified SBPs (mol). The mixture was incubated at 30 °C for 15 min. After incubation, the product was purified on a Sephadex G-25 column equilibrated with buffer B. The MAL-R-mixed surface-modified SBPs were collected and aliquoted at -40 °C in the same buffer.

[0238] Example 15 Mixed surface-modified SBP-antibody conjugates With stirring, MAL﹣R﹣mix﹣m﹣SB﹣PPI﹣64﹣NH2 / OH﹣2 (154 μL, 16.6 nmol) was added to the reduced antibody (14 nmol, in 220 μL). The pH was adjusted to approximately 6.8 by adding 12.5 μL of sodium carbonate (1.0 M solution), the reaction was continued at room temperature for 1 h, and terminated by adding 100 μL of cysteine ​​(0.4 mM solution). The conjugated mixture was purified by elution with a sodium chloride gradient on a CM cellulose column.

[0239] Example 16 IgG-asymmetric random branched polymer conjugate The preparation of randomly branched mixed surfaces (OH / NH2 mix) m﹣ran﹣AB﹣PEI﹣NH2 / OH﹣2﹣IgG conjugates is provided as a general procedure for preparing polymer-antibody conjugates.

[0240] Other conjugates, such as PEI﹣IgG, m﹣ran﹣AB﹣PEI﹣NH2﹣1﹣IgG, m﹣ran﹣AB﹣PEI﹣NH2﹣2﹣IgG, m﹣ran﹣AB﹣PEI﹣NH2﹣3﹣IgG, m﹣ran﹣AB﹣PEI﹣NH2﹣4﹣IgG, and m﹣ran﹣AB﹣PEI﹣NH2 / OH﹣1(OH / NH2mix)﹣IgG, m﹣ran﹣AB﹣PEI﹣NH2 / OH﹣2(OH / NH2 Mix)﹣IgG, m﹣ran﹣AB﹣PEI﹣NH2 / OH﹣3(OH / NH)2mix)﹣IgG, regular polylysine polymers, and randomly branched poly(2-ethyloxazoline) with alkyl-terminal primary amine chains were all synthesized in a similar manner. The synthesis of various protein conjugates of asymmetrically randomly branched PEOX polymers was also carried out in a similar manner.

[0241] Example 17 LC﹣SPDP﹣Mixed Surface m﹣ran﹣AB﹣PEI﹣NH 2 / OH﹣2 The randomized m﹣ran﹣AB﹣PEI﹣NH2 / OH﹣2 (4×10⁻⁶) surface-branched molecules were introduced into 400 μL of phosphate buffer (20 mM phosphate and 0.1 M NaCl, pH 7.5). ﹣7 4 × 10 mol) added to 400 μl of water ﹣6 1 mol of sulfonyl-LC-SPDP (Pierce, IL) was vortexed and incubated at 30 °C for 30 min. LC-SPDP-m-ran-AB-PEI-NH2 / OH-2 was purified by gel filtration chromatography and equilibrated with buffer A (0.1 M phosphate, 0.1 M NaCl, and 5 mM EDTA, pH 6.8). The product was further concentrated to obtain 465 μl of a solution with a concentration of approximately 0.77 nmol.

[0242] Example 18 Thiolized m﹣ran﹣AB﹣PEI﹣NH 2 / OH﹣2 LC-SPDP m-ran-AB-PEI-NH2 / OH-2 (50 nmol in 65 mL of buffer A) was mixed with 100 μL of dithiothreitol (DTT) (50 mM in buffer A) and allowed to incubate at room temperature for 15 minutes. Excess DTT and byproducts were removed by gel filtration through buffer A. The product was concentrated in a 10 K Centricon concentrator to obtain 390 μL of thiolized m-ran-AB-PEI-NH2 / OH-2, which was used for conjugation with activated antibodies.

[0243] The maleimide-R-activated antibody, as described above, is used for conjugation with thiolized m-ran-AB-PEI-NH2 / OH-2.

[0244] Example 19 m﹣ran﹣AB﹣PEI﹣NH 2 / OH﹣2﹣antibody conjugate Add MAL-R activated antibody (4.8 mL or 34 nmol) to the thiolized m-ran-AB-PEI-NH2 / OH-2 (310 μL or 35.7 nmol). Concentrate the reaction mixture to approximately 800 μL and allow it to incubate overnight at 4°C and / or for approximately 1 hour at room temperature. After completion, quench the reaction with 100 μL of ethylmaleimide (50 mM solution), and then fractionate the conjugate on a CM cellulose column (5 mL) using a sodium chloride gradient in 20 mM phosphate buffer at pH 6. The conjugate was eluted with a sodium chloride gradient and characterized by cation exchange chromatography, UV spectroscopy, and polyacrylamide gel electrophoresis.

[0245] Example 20 Preparation of paclitaxel formulations and nanoparticles As a standard procedure, paclitaxel is dissolved in methanol to a concentration up to 40 mg / mL. C 18 The -PEOXABP60 polymer was dissolved alone in methanol to concentrations up to 100 mg / mL. The two solutions were then mixed in different volumes to achieve a final polymer-to-paclitaxel weight ratio in the mixture ranging from 3:1 to 10:1. The mixture was then lyophilized for 20 to 96 hours, depending on the volume.

[0246] For example, the diameter of the aggregates, as measured by light scattering, was approximately 70 nm to 90 nm before freeze-drying and 120 nm to 140 nm after freeze-drying.

[0247] Alternatively, paclitaxel and C 18Both -PEOXABP60 polymers are soluble in common solvents such as acetone, methanol, or ethanol, and are then added dropwise to water while being stirred or sonicated, followed by sterile filtration through a 0.22 μm filter. The final product can then be obtained by lyophilization, and the size of the aggregates can be measured by light scattering.

[0248] Other taxane-induced aggregates or nanoparticles of branched polymers with various hydrophobic surface modifications, such as C4, C6, and C4, can be prepared in a similar manner. 12 Or C 22 Hydrocarbon-modified random branched PEOX, PEI, and PPI polymers: C4, C6, C 12 C 18 and C 22 Hydrocarbon-modified PAMAM, PEI, and PPI dendritic polymers and dendritic grafts; and C4, C6, C 12 C 18 and C 22 Hydrocarbon-modified branched PLL / polymer.

[0249] Example 21 With 7:1 C 18 -PEOXABP60 polymer:paclitaxel ratio nanoparticles Paclitaxel (700 mg) was dissolved in 9.33 mL of methanol to obtain a 75 mg / mL solution. A 15 mg / mL paclitaxel solution was also prepared by dissolving 100 mg of paclitaxel in 6.67 mL of methanol. The two solutions were mixed for 20 minutes to obtain a solution containing 6.25 mg of paclitaxel and 43.75 mg of polymer per mL, providing a polymer:drug ratio of 7:1. The mixture was placed on a rotary evaporator and the methanol was removed until dry. The resulting solid was redissolved in 33.3 mL of water with stirring to obtain a final paclitaxel concentration of 3 mg / mL. The solution was passed through a 0.8 μm filter and then through a 0.22 μm filter. Depending on the volume used, the filtrate was lyophilized over a period of 24–72 hours. The vial was sealed, and the ready-to-use white powder was stored at room temperature. This product is designated as FID-007.

[0250] Example 22 With C 18 -PEOXABP80 and C 18 -PEOXABP50 and paclitaxel nanoparticles Polymer C 18 -PEOXABP80 and C 18The -PEOXABP50 was used to produce polymer-drug at a polymer:drug ratio of 7:1 according to the same procedure described above. Particle size was measured and is shown in Table 1 below.

[0251] Example 23 Comparison of polymer C 18 -PEOXABP200 and C 18 -PEOXABP100 and paclitaxel nanoparticles Using the same procedure, two comparative polymers C with different monomer / initiator ratios were compared. 18 -PEOXABP200 and C 18 -PEOXABP100 was used to generate comparative aggregates at two different polymer:drug weight ratios of 5:1 and 7:1. Aggregate sizes were measured, and the data are shown in Table 1 below.

[0252] Nanoparticle Measurement The sizes of various polymers, polymer-only aggregates, and drug-induced polymer aggregates were measured using a Malvern Zetasizer Nano-ZS Zen3600 particle size analyzer (Malvern Panalytical Inc., Westborough, MA) via dynamic light scattering.

[0253] As shown in Table 1, polymers with a monomer-to-initiator molar ratio of 100:1 or higher produce polymer-drug aggregates with a polymer:drug weight ratio of 5:1 and 7:1, resulting in aggregates with a size exceeding 100 nm. Polymers with a monomer-to-initiator molar ratio of 80:1, 60:1, 50:1, or 20:1 produce polymer-drug aggregates with a particle size range of approximately 70 nm to 100 nm.

[0254] The value of D(v,0.9) (also known as D) 90 The particle size distribution can be obtained using, for example, a Malvern Zetasizer Nano-ZS Zen3600 particle size analyzer. The D... 90 The value is the value of the number of particles that have a size (in diameter) that is below, smaller than, or lower than that value. (By C) 18 -PEOXABP100 (comparison) and C 18 The polymer-drug aggregate D prepared by -PEOXABP60 (an embodiment of this disclosure) 90 The values ​​are shown in Table 1.

[0255] Table 1. Particle size of polymer-drug aggregates. Filtration rate measurement The R-values ​​of the polymer-drug aggregates prepared above were measured according to the following procedure. f Samples of each polymer-drug aggregate were prepared and dissolved in water to the same final concentration as described herein. A final paclitaxel concentration of 3 mg / mL was used for the samples, and the data are presented in Table 2 below.

[0256] Load each sample at the starting volume (V0) into a sterile syringe. Attach a disposable 25 mm sterile 0.22 μm syringe filter assembly (Pall Corp., Ann Arbor, MI) to the syringe. Use a 9 ml V0 volume per sample. For the 25 mm filter, the starting volume can range from 9 ml to 10 ml.

[0257] Then, each sample is passed through a 0.22 μm filter by pressing the plunger of the syringe with constant thumb pressure until all sample volumes have passed through the filter or the plunger no longer moves forward under the same pressure.

[0258] Record the remaining sample volume (V1) in the syringe. Based on formula V f =V0﹣V1 Calculate the filtration volume (V f ) or filtrate. Optionally, the sample passing through the filter can be collected, and V f The volume of the filtrate or the collected filtered sample is measured.

[0259] Based on equation R f =V f / V0 calculates R for each sample. f .

[0260] For C 18 Polymer-drug aggregates prepared from PEOXABP60 and paclitaxel, all 9 ml, were passed through a filter. f =9, thus obtaining R f =1 or 100%.

[0261] Comparison of polymer-drug aggregates by C 18 Preparation of PEOXABP100 and paclitaxel. Only 4.4 ml passed through the filter. Therefore, V f =4.4, thus obtaining R f =4.4 / 9 or 0.489 or 48.9%. The data is shown in Table 2.

[0262] Table 2. Filtration rate of polymer-drug aggregates. Activity test Metabolism in living cells produces "reducing equivalents," such as NADH or NADPH. These reducing compounds transfer electrons to an intermediate electron transfer reagent, which reduces the tetrazolium product MTS (Promega) to a colored, water-soluble formazanine product. Upon death, cells rapidly lose the ability to reduce the tetrazolium product. Therefore, the production of the colored formazanine product is proportional to the number of living cells in the culture.

[0263] CellTiter 96 ® AQ ueous The product (Promega, Madison, WI) is a material and method for performing MTS assays to determine the number of viable cells in a culture. A single reagent is added directly to the assay well at the recommended ratio of 20 μl reagent to 100 μl culture medium. Cells are incubated at 37°C for 1-4 hours, and then absorbance is measured at 490 nm.

[0264] Example 24 Toxicity and potency of nano-encapsulated paclitaxel / ABP60 (FIB-007) As previously described, using C 18 Nanoparticles of paclitaxel were prepared using the PEOXABP60 polymer at a polymer-to-paclitaxel ratio of 7:1. In cytotoxicity studies using normal human skin fibroblast cell lines and various cancer cell lines, as well as in vivo studies of toxicity (maximum tolerated dose, MTD) and inhibition of tumor growth in three mouse xenograft models, this formulation, named FID-007, was compared with TAXOL and ABRAXANE.

[0265] Example 25 In vitro activity of FID-007 In in vitro cytotoxicity assays, FID-007 was tested with TAXOL and ABRAXANE on normal human fibroblasts and various cancer cell lines. Although FID-007 inhibited the in vitro proliferation of a range of human cancer cell lines, including those derived from breast cancer, ovarian cancer, and lung cancer cells, it exhibited low cytotoxicity against normal cells, similar to levels observed with TAXOL and ABRAXANE. Figure 15 Overall, FID-007 showed 10 times lower toxicity to normal cells than to tumor cells, exhibiting very high EC50 greater than 100 μM. 50 (Drug concentration at which half-maximal response is achieved). FID-007 was active in a 72-h toxicity assay in the human lung cancer cell line A549, EC50. 50 2.8 ng / mL Figure 16 FID-007 is cytotoxic to MDA-MB-231 (triple-negative breast cancer cells), EC...50 It was 4.9 ng / mL. Figure 17 FID-007 is cytotoxic to OV-90 (ovarian cancer cells), EC... 50 5.0 ng / mL Figure 18 For all three cancer cell lines, FID-007 showed comparable cytotoxicity to TAXOL and ABRAXANE.

[0266] Example 26 In vivo activity of FID-007 A series of experiments were conducted to determine the in vivo tolerability, activity, and basic pharmacokinetics of intravenously (IV) administered FID-007 in mice compared to TAXOL and ABRAXANE. FID-007 was well tolerated at daily doses up to 150 mg / kg. To confirm its antitumor activity, FID-007 was IV administered daily to mice at well-tolerated doses in three different mouse xenograft models (lung, ovarian, and breast cancer). Generally, FID-007 was better tolerated in mouse xenograft models than standard cytotoxic agents with similar targets (e.g., TAXOL and ABRAXANE) and selectively inhibited tumor growth.

[0267] The half-life of FID-007 in mice was determined to be approximately 9.3 hours using an optimized HPLC method. The liver and spleen, followed by blood, were the organs with the highest FID-007 concentrations at 1 hour. PK profiles of FID-007, TAXOL, and ABRAXANE are shown in [Figure / Table / Insert Table ... Figure 19 middle.

[0268] In one study, the single-dose MTD of FID-007 was compared with that of TAXOL and ABRAXANE, in which different doses of the drug were administered via the tail vein of healthy CD1 mice and SCID (immunodeficient) mice over a period of several weeks. Control mice were administered saline. The single-dose MTDs of TAXOL, ABRAXANE, and FID-007 in CD1 mice were found to be 20 mg / kg, 240 mg / kg, and 175 mg / kg, respectively. No major side effects were observed in all surviving mice. However, weight gain was observed in all treatment groups for ABRAXANE and FID-007 compared to the control group (treated with saline). ABRAXANE at doses of 120 mg / kg and above caused a dose-dependent increase in weight. The same results were observed with FID-007 at doses of 150 mg / kg and higher.

[0269] The multidose MTD of FID-007 was similarly determined by administering FID-007 (100 mg / kg and 150 mg / kg) via tail vein to healthy CD-1 mice and SCID mice (10 weeks old, female) on days 0, 3, and 6. Animals were monitored twice daily and weighed every 3 days. The multidose MTD of FID-007 in CD-1 mice was determined to be 100 mg / kg, and in SCID mice it was determined to be 30 mg / kg, with some side effects immediately following injection. There was no excessive weight gain in the FID-007 multidose groups compared to the control group.

[0270] In mouse models of human lung, breast, and ovarian cancer xenografts, the in vivo efficacy of FID-007 in inhibiting tumor growth was compared with that of TAXOL and ABRAXANE. Sixty female and male SCID mice (6-8 weeks, 20-26 g, Charles River; 40 females for breast and ovarian cancer, and 20 males for lung cancer) were injected with 0.1 mL of serum-free medium containing a suspension of lung cells A549, breast cells MDA-MB-231, or ovarian cells OV90. Cells were pre-cultured in a humidified incubator (37°C, 5% CO2, 95% air). Each mouse tumor was treated with 3 × 10⁶ cells. 6 (A549), 10 7 (MDA﹣MB﹣231) and 5×10 6 (OV-90) cell dose. Tumor growth was allowed to continue for 7 to 9 days prior to treatment initiation, and all tumor volume measurements were obtained using digital calipers (VWR Inc.). (W...) 2 The tumor volume was calculated as (W × L) / 2, where W is the maximum tumor width and L is the maximum tumor length. Tumor and body weight measurements were obtained on the same day before the first treatment, and then every three days thereafter. Day 0 was designated as the first day of treatment. On day 0, animals developing tumors were randomly assigned to five groups [approximately 4 mice per group (8 tumors)], with each treatment group representing a wide range of tumor sizes. The volume of the OV-90 xenograft tumor was... Figure 22 As shown in the image.

[0271] ABRAXANE (80 mg / kg), FID-007 (20 mg / kg), TAXOL (20 mg / kg) and C 18The ABP60 polymer starting material, designated NanoCarrier-001B (20 mg / kg), was freshly prepared for each injection. Saline was used as a mediator control. The drug or saline was administered via tail vein injection every three days. Drug doses were selected to ensure isotoxicity across all treatment groups based on previously determined single-dose and multi-dose MTDs. The lung cancer, breast cancer, and ovarian cancer groups each received a total of four injections. Throughout the study, the injection volume for controls, ABRAXANE, and FID-007 was 0.1 mL per injection. Due to the viscosity of the TAXOL formulation, 0.2 mL per injection was administered for a dose of 20 mg / kg. Mean body weight and tumor volume measurements were calculated by averaging the results across animals within the same group. Mice were euthanized with isoflurane 21 days after the last treatment for lung and ovarian cancer, and 10 days after the last treatment for breast cancer. Blood and isolated serum, as well as tumor tissue and liver, were collected and stored at -80°C.

[0272] In the lung cancer (A549) xenograft group, overall, no deaths occurred in any treatment group. Possibly due to TAXOL toxicity, heavy breathing and inactivity were observed in several mice within the first 30 minutes after treatment. Mean body weight and tumor volume measurements were calculated by averaging within the same group. The overall mean body weight increases for saline control, TAXOL, FID-007, and nanocarrier control were 6.05%, 5.87%, 6.38%, and 12.3%, respectively. However, all mice in the ABRAXANE group developed neurotoxicity and lost >20% of their body weight. These mice were sacrificed on day 13. For the saline control group, the tumor volume increased by 1827 mm. 3 Furthermore, in the NanoCarrier-001B mediator control group, tumor volume increased by 1311 mm. 3 In the TAXOL group, the tumor volume increased by 305.8 mm. 3 However, the tumor volume in the FID-007 group decreased by 39.7 mm. 3 ( Figure 20 ). Figure 21 and Figure 23 Representative images of tumors in the treatment group are shown.

[0273] No deaths occurred in the breast cancer (MDA-MB-231) xenograft group in any treatment group. Heavy breathing and inactivity were observed within the first 30 minutes after treatment, possibly due to TAXOL toxicity. In the ABRAXANE group, all mice exhibited hind leg weakness and a 20% weight loss after three treatments, leading to the decision to discontinue a fourth treatment in this group. Mean body weight and tumor volume measurements were calculated by averaging within the same group. The overall mean weight gain for saline, TAXOL, FID-007, and NanoCarrier-001B was 3.76%, 0.46%, 1.8%, and 4.2%, respectively. For the ABRAXANE group, the mean weight loss was 7.66%. Tumor volume increased by 328.6 mm in both the saline and NanoCarrier-001B groups. 3 and 458.8 mm 3 In the FID-007, TAXOL, and ABRAXANE groups, tumor volume decreased by 108.7 mm. 3 75.5 mm 3 and 70.2 mm 3 Tumor volume observation results are shown in Figure 23 middle.

[0274] In the ovarian cancer (OV-90) xenograft group, the TAXOL treatment group showed some toxicity, with heavy breathing and inactivity observed in two mice within the first 30 minutes after treatment. The mean weight gain was 3.23%, 17.1%, 13.5%, 15.4%, and 2.24% in the saline control, TAXOL, ABRAXANE, FID-007, and NanoCarrier-001B control groups, respectively. Tumor volume increased by 652.7 mm in the saline control, NanoCarrier-001B control, and TAXOL groups, respectively. 3 271.9 mm 3 and 9.1 mm 3 In the FID-007 group, the tumor volume decreased by 93.1 mm. 3 In the ABRAXANE group (80 mg / kg), tumor volume decreased by 72.4 mm. 3 .

[0275] FID-007 demonstrated in vitro cytotoxicity against lung, breast, and ovarian cell lines similar to established antitumor drugs TAXOL and ABRAXANE, while maintaining low levels of toxicity to normal cells. In mouse xenograft models of human lung, breast, and ovarian cancer, FID-007 showed in vivo efficacy comparable to or significantly better than the two approved drugs in inhibiting tumor growth and reducing tumor mass.

[0276] Example 27 Forty-six patients participated in the Phase I study, including 11 patients with HNSCC. Primary tumor sites included two nasopharyngeal tumors, two nasal cavity and paranasal sinuses, five oropharyngeal tumors, one oral cavity tumor, and one occult primary tumor. FID-007 was administered at 15 mg / m² on days 1, 8, and 15 of a 28-day cycle. 2 Up to 160 mg / m 2 The dose level was administered IV. The median age was 61 years (53–75). The ECOG PS was 0–2 in all 40 patients. The median prior therapy was 3 (1–5), and all had received prior immune checkpoint inhibitors. Seven patients had previously received taxane chemotherapy. Among all solid tumors, the partial response (PR) rate was 17% (8 patients), the stable disease (SD) rate was 35% (16 patients), and the progressive disease (PD) rate was 46% (21 patients). In the HNSCC group of 11 patients, 5 patients (46%) had a partial response (PR), 3 patients (27%) had a stable disease (SD), and 3 patients (27%) had progressive disease (PD). Three of the five HNSCC patients with a partial response (PR) had previously been treated with taxane. The overall response rate (ORR) for all treated HNSCC patients was 45%. The treatment duration was 4 months, ranging from 1 month to 15 months. FID-007 was administered intravenously over 30–60 minutes, once weekly for 3 weeks of a 28-day treatment cycle. Sodium bicarbonate infusions (before and after treatment) were used for prophylaxis at a dose of 15 mg / m². 2 Up to 125 mg / m 2 Potential nephrotoxicity in patients at dose levels of FID-007. Sodium bicarbonate infusion solution contains saline diluent (0.9% sodium chloride injection, USP) and 0.15% sodium bicarbonate (USP).

[0277] Examples of representative images of tumor size in Figure 24A - Figure 24B As shown in the figure. In one patient, the size of the tumor (1) before treatment was approximately 28 mm. Figure 24AAfter two cycles of treatment with FID-007, the size of tumor (2) was approximately 15.30 mm. The patient had prior therapy (best response): pembrolizumab + 5-FU+, carboplatin (SD), cetuximab (SD), docetaxel (PR 9 months), NK cells + EGFR bispecific Ab (PD), with a response lasting >6 months.

[0278] Example 28 Typically, during a phase II study, eligible patients are recruited and randomly (1:1) assigned to FID-007 and a fixed dose of cetuximab (500 mg / m² every two weeks, starting on day 1 and day 15 of each 28-day cycle, beginning treatment cycle 2). 2 One of two groups for intravenous (IV) infusion: Group A: FID-007 (75 mg / m²) 2 Add cetuximab, or group B: FID-007 (125 mg / m²) 2 In addition to cetuximab, patients received FID-007 via IV infusion at the designated dose over 30 (±5) minutes on days 1, 8, and 15 of each 28-day cycle. Cetuximab was administered after the completion of the FID-007 infusion, starting on day 1 of cycle 2. Tumor response was assessed every 8 weeks (i.e., every 2 cycles of study drug treatment) according to RECIST version 1.1 after screening and initiation of cycle 3. Patients continued to receive cetuximab and FID-007 until they met the study drug discontinuation criteria. There was no maximum limit to the number of treatment cycles a patient could receive. Randomization was stratified by p16 status (positive or negative) and prior taxane exposure. Patients were monitored and evaluated by qualified physicians.

[0279] Patients participated in the study to receive FID-007 and ERBITUX ® Treatment with a combination of cetuximab and other medications. As described above, each patient was administered 75 mg / m² via intravenous (IV) injection. 2 FID-007 plus cetuximab (starting from treatment cycle 2, 500 mg / m² every 2 weeks on days 1, 8, and 15 of each 28-day cycle). 2 Intravenous (IV) infusion. Patients are monitored and evaluated by a qualified physician.

[0280] Patients participated in the study to receive FID-007 and ERBITUX ® Treatment with a combination of cetuximab and other medications. As described above, each patient was administered 125 mg / m² via intravenous (IV) injection. 2 FID-007 plus 500 mg / m² in the same cycle 2Cetuximab was administered intravenously (IV). Patients were monitored and evaluated by qualified physicians. Patients enrolled in the study were those with recurrent or metastatic HNSCC whose disease had progressed after treatment with a programmed death-ligand 1 (PD-L1)-based immune checkpoint inhibitor and ≤1 line of anticancer therapy for recurrent / metastatic disease. Eligibility was reviewed and confirmed during a 28-day screening period prior to enrollment. During screening, their p16 protein expression (in oropharyngeal carcinoma only) and PD-L1 expression were tested.

[0281] Receive 125 mg / m² via intravenous (IV) administration. 2 FID-007 plus 500 mg / m 2 Infusion reactions were observed in some patients receiving intravenous (IV) cetuximab. Following the first treatment cycle, they were given a reduced dose of FID-007 at a treatment cycle of 100 mg / m² via intravenous (IV) injection. 2 FID-007 plus cetuximab (starting from treatment cycle 2, 500 mg / m² every 2 weeks on days 1, 8, and 15 of each 28-day cycle). 2 The combination of intravenous (IV) infusions, as described above, should be monitored and evaluated by a qualified physician.

[0282] Example 29 In patients recruited for the Phase II study where tumor assessment results were received: the FID-007 dose level was 125 mg / m². 2 And 125 mg / m 2 Then reduce to 100 mg / m³ 2 Some patients have achieved a complete response (CR), which is the disappearance of all signs of cancer in response to treatment, also known as complete remission; the FID-007 dose level is 75 mg / m². 2 and 125 mg / m 2 Then 125mg / m 2 Reduced to 100 mg / m 2 Some patients have a partial response (PR), which is a reduction in tumor size or degree of cancer in response to treatment; also known as partial remission; and the FID-007 dose level is 75 mg / m². 2 and 125 mg / m 2 Then 125 mg / m 2 Reduced to 100 mg / m 2 Some patients have stable disease (SD), meaning that the degree or severity of the cancer neither decreases nor increases.

[0283] All references cited in this article are incorporated into this article in their entirety through citation.

[0284] It should be understood that various changes and modifications may be made to the teachings herein without departing from the spirit and scope of this disclosure.

Claims

1. A method for treating cancer in a subject with a corresponding need, the method comprising administering an effective dose of a first pharmaceutical composition to the subject, wherein the first pharmaceutical composition comprises an aggregate, the aggregate comprising: (a) A polyoxazoline comprising at least one first end group modified by a hydrophobic moiety, wherein the polyoxazoline further comprises a linear moiety, a branched moiety, or both; the branched moiety comprises a symmetrically branched polymer, an asymmetricly branched polymer, or a combination thereof; and the polyoxazoline comprises a monomer to initiator molar ratio in the range of 50:1 to 80:1; and (b) Taxane, and The subject in question has recurrent or metastatic cancer and has been treated with radiation therapy, surgery, chemotherapy, hormone therapy, biotherapy, immunotherapy, checkpoint inhibitor therapy, programmed death-ligand 1 (PD-L1)-based immune checkpoint inhibitor therapy, or a combination thereof, for at least 7 days prior to the first administration of the first drug composition.

2. The method of claim 1, wherein the aggregate comprises a weight ratio of the polyoxazoline to taxane of 6:1 to 8:1; and has a size of 70 nm to 90 nm.

3. The method according to claim 1, wherein the polyoxazoline comprises a second end group containing a functional group modified with ethylenediamine (EDA) or an EDA derivative, wherein the ratio of the second end group to EDA is 1:

10.

4. The method of claim 3, wherein the EDA derivative is configured to include diethylenetriamine, triethylenetetramine, tetraethylenepentamine, pentaethylenehexamine, polyvinylamine, or tetramethylethylenediamine.

5. The method of claim 1, wherein the subject has been treated with at least one drug, said drug comprising afatinib, albumin-bound paclitaxel or nab-paclitaxel, bleomycin sulfate, carboplatin, cisplatin, cetuximab, docetaxel, cetuximab, Hydrea (hydroxyurea), hydroxyurea, pembrolizumab, methotrexate, nivolumab, cisplatin, paclitaxel, docetaxel, Trexall (methhotrexate), a combination of carboplatin and Taxol, a combination of TPF (docetaxel, cisplatin, and fluorouracil (FU)), or any of the foregoing combinations.

6. The method of claim 1, wherein the subject has been treated with: pembrolizumab as a single agent with a combination positive score (CPS) ≥1; a combination of pembrolizumab, platinum, and fluorouracil; cetuximab; docetaxel; nivolumab; afatinib; or any combination of the foregoing.

7. The method of claim 6, wherein the subject has been treated with: first-line treatment comprising the pembrolizumab or a combination of the pembrolizumab, platinum, and fluorouracil; and second-line treatment comprising cetuximab, docetaxel, nivolumab, afatinib, or a combination thereof.

8. The method according to claim 1, wherein the cancer is breast cancer, triple-negative breast cancer, ovarian cancer, lung cancer, NSCLC (non-small cell lung cancer), colon cancer, gastric cancer, melanoma, head and neck cancer (HNC), pancreatic cancer, or a combination thereof.

9. The method according to claim 8, wherein the head and neck cancer (HNC) is squamous cell carcinoma of the head and neck (SCCHN), unresectable HNC, squamous cell carcinoma of the head and neck (HNSCC), human papillomavirus (HPV) positive head and neck cancer, metastatic HNC, metastatic HNC with three or more metastatic sites, occult primary metastatic squamous cell carcinoma, pharyngeal cancer, hypopharyngeal cancer, laryngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, oral cancer, sinus cancer, nasal cavity sinus cancer, nasal cavity cancer, salivary gland cancer, thyroid cancer, recurrent HNC, refractory HNC, HNC resistant to PD-1 therapy, HNC resistant to PD-L1 therapy, HNC resistant to immunotherapy, HNC resistant to T-cell therapy, HNC resistant to CAR-T therapy, HNC resistant to NK cell therapy, or a combination thereof.

10. The method of claim 1, wherein the initiator is configured to comprise a hydrophobic electrophilic molecule.

11. The method of claim 1, wherein the initiator is configured to comprise a hydrocarbon.

12. The method of claim 11, wherein the hydrocarbon is configured to contain 1 to 22 carbons, and the hydrocarbon may be saturated or unsaturated.

13. The method of claim 1, wherein the initiator is configured to include an aliphatic hydrocarbon, an aromatic hydrocarbon, or a combination of both.

14. The method of claim 1, wherein the initiator is configured to contain a halide functional group.

15. The method of claim 1, wherein the initiator is configured to include alkyl halides, aralkyl halides, acyl halides, or combinations thereof.

16. The method of claim 1, wherein the initiator is configured to include methyl iodine, methyl bromide, methyl chloride, ethyl iodine, ethyl bromide, ethyl chloride, 1-iodopropane, 1-bromopropane, 1-chloropropane, 1-iodobutane, 1-bromobutane, 1-chlorobutane, 1-iopentane, 1-bromopentane, 1-chloropentane, 1-iohexane, 1-bromohexane, 1-chlorohexane, 1-iodododecane, 1-bromododecane, 1-chlorododecane, 1-iooctadecane, 1-bromooctadecane, 1-chlorooctadecane, benzyl iodine, benzyl bromide, benzyl chloride, allyl bromide, acyl iodine, acyl bromide, acyl chloride, benzoyl bromide, benzoyl chloride, or combinations thereof.

17. The method of claim 1, wherein the initiator is configured to contain a p-toluenesulfonyl group.

18. The method of claim 1, wherein the taxane is configured to associate with the at least one first end group.

19. The method of claim 1, wherein the polyoxazoline is configured to include poly(2-oxazoline), poly(2-substituted oxazoline), or a combination thereof.

20. The method of claim 1, wherein the polyoxazoline is configured to include poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-butyloxazoline), or combinations thereof.

21. The method of claim 1, wherein the taxane is configured to include paclitaxel, docetaxel, or a combination thereof.

22. The method of claim 1, wherein the aggregate comprises a polyoxazoline to taxane ratio of 7:

1.

23. The method of claim 1, wherein the first pharmaceutical composition is administered to the subject via parenteral administration, the parenteral administration being selected from intravenous (IV) injection, intradermal injection, subcutaneous injection, or a combination thereof.

24. The method of claim 1, further comprising the step of administering an effective dose of a second pharmaceutical composition to the subject simultaneously or sequentially with the first pharmaceutical composition, the second pharmaceutical composition comprising carboplatin, cisplatin, a monoclonal antibody or fragment thereof that binds to EGFR, a bispecific antibody or fragment thereof having at least one binding site for binding to EGFR, a trivalent antibody or fragment thereof having at least one binding site for binding to EGFR, a small molecule drug that binds to EGFR, or a combination thereof.

25. The method of claim 24, wherein the first pharmaceutical composition and the second pharmaceutical composition are formulated as a single-dose unit comprising the first pharmaceutical composition and the second pharmaceutical composition, the single-dose unit being selected from injectable packages containing the first pharmaceutical composition and the second pharmaceutical composition separately in a common package, premixed injectable packages comprising a mixture of the first pharmaceutical composition and the second pharmaceutical composition, or combinations thereof.

26. The method of claim 24, wherein the EGFR-binding monoclonal antibody or a fragment thereof is cetuximab.

27. The method of claim 24, wherein the effective dose of the first pharmaceutical composition comprises 15 mg / m². 2 Up to 500 mg / m 2 The taxane is within the range described above, and the effective dose of the second pharmaceutical composition comprises 50 mg / m². 2 Up to 900 mg / m 2 The cetuximab mentioned above is within the range of [specific range].

28. The method of claim 24, wherein the effective dose of the first pharmaceutical composition and the effective dose of the second pharmaceutical composition are administered to the subject at least once on day 1, day 8, day 15, day 28, or a combination thereof during a treatment cycle.

29. The method of claim 1, wherein the aggregate is configured to further comprise a targeting portion.

30. The method of claim 29, wherein the targeting portion is configured to include an antibody or a fragment thereof, its antigen-binding portion, an antigen, a cell surface receptor, a cytoplasmic receptor, a cell receptor ligand, or a lectin ligand.

31. A pharmaceutical composition comprising: A first pharmaceutical composition comprising an aggregate, said aggregate comprising: (a) A polyoxazoline comprising at least one first end group modified by a hydrophobic moiety, wherein the polyoxazoline further comprises a linear moiety, a branched moiety, or both, the branched moiety comprising an asymmetricly branched polymer; and the polyoxazoline comprises a monomer to initiator molar ratio of 60:

1. (b) Taxane, The aggregate comprises a polyoxazoline to taxane weight ratio of 6:1 to 8:1; and has a size of 70 nm to 90 nm; and the polyoxazoline comprises a second end group containing a functional group modified with ethylenediamine (EDA) or an EDA derivative, wherein the ratio of the second end group to EDA is 1:10; and The second pharmaceutical composition comprises carboplatin, cisplatin, a monoclonal antibody or fragment thereof that binds to EGFR, a bispecific antibody or fragment thereof having at least one binding site for binding to EGFR, a trivalent antibody or fragment thereof having at least one binding site for binding to EGFR, a small molecule drug that binds to EGFR, or a combination thereof. The pharmaceutical composition is formulated as a single-dose unit comprising the first pharmaceutical composition and the second pharmaceutical composition containing the taxane, the single-dose unit being selected from injectable packages containing the first pharmaceutical composition and the second pharmaceutical composition separately in a common package, premixed injectable packages containing a mixture of the first pharmaceutical composition and the second pharmaceutical composition, or combinations thereof.

32. The pharmaceutical composition of claim 31, wherein the EGFR-binding monoclonal antibody or a fragment thereof is cetuximab.

33. The pharmaceutical composition of claim 31 further comprises a salt component comprising sodium bicarbonate, and wherein the pH of the pharmaceutical composition is in the range of 7.1 to 10.

34. Use of a first pharmaceutical composition and a second pharmaceutical composition for manufacturing a medicament for treating head and neck cancer in a subject with a corresponding need, wherein the first pharmaceutical composition comprises an aggregate comprising: (a) A polyoxazoline comprising at least one first end group modified by a hydrophobic moiety, wherein the polyoxazoline further comprises a linear moiety, a branched moiety, or both, the branched moiety comprising an asymmetricly branched polymer; and the polyoxazoline comprises a monomer to initiator molar ratio of 60:

1. (b) Taxane, The aggregate comprises a polyoxazoline to taxane weight ratio of 6:1 to 8:1; and has a size of 70 nm to 90 nm; and the polyoxazoline comprises a second end group containing a functional group modified with ethylenediamine (EDA) or an EDA derivative, wherein the ratio of the second end group to EDA is 1:10; and The second pharmaceutical composition comprises carboplatin, cisplatin, a monoclonal antibody or fragment thereof that binds to EGFR, a bispecific antibody or fragment thereof having at least one binding site for binding to EGFR, a trivalent antibody or fragment thereof having at least one binding site for binding to EGFR, a small molecule drug that binds to EGFR, or a combination thereof. The drug is formulated as a single-dose unit comprising the first drug composition and the second drug composition, the single-dose unit being selected from injectable packages containing the first drug composition and the second drug composition separately in a common package, premixed injectable packages containing a mixture of the first drug composition and the second drug composition, or combinations thereof.

35. The use according to claim 34, wherein the EGFR-binding monoclonal antibody or a fragment thereof is cetuximab.

36. The use according to claim 34, wherein an effective dose of the first pharmaceutical composition and an effective dose of the second pharmaceutical composition are administered to the subject at least once on day 1, day 8, day 15, day 28, or a combination thereof during a treatment cycle.

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