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WO2016000070A1 - Polyglycérol hyperramifié à dérivatisation hydrophobe pour l'administration d'un médicament par voie intravasculaire - Google Patents

Polyglycérol hyperramifié à dérivatisation hydrophobe pour l'administration d'un médicament par voie intravasculaire Download PDF

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Publication number
WO2016000070A1
WO2016000070A1 PCT/CA2015/050600 CA2015050600W WO2016000070A1 WO 2016000070 A1 WO2016000070 A1 WO 2016000070A1 CA 2015050600 W CA2015050600 W CA 2015050600W WO 2016000070 A1 WO2016000070 A1 WO 2016000070A1
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Prior art keywords
hyperbranched polyglycerol
nanoparticle
hpg
core
alkyl chains
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English (en)
Inventor
Mohamed K. Khan
Jayachandran N. Kizhakkedathu
Ripen Misri
Rajesh A. SHENOI
Nelson K. Y. WONG
Donald E. Brooks
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University of British Columbia
British Columbia Cancer Agency BCCA
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University of British Columbia
British Columbia Cancer Agency BCCA
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner

Definitions

  • This invention relates to therapeutics, their uses and methods for drug delivery.
  • the invention relates to polymers based on derivatized hyperbranched polyglycerols and methods for intravascular delivery of biologically active moieties to biological tissues.
  • DTX docetaxel
  • Tween 80 polysorbate 80
  • Nanoparticles can improve the solubility of poorly water-soluble drugs, extend the circulation half- life of the drug in plasma, and provide controlled drug release that can result in enhanced therapeutic benefits in cancer treatment. Accumulation of NPs in solid tumors is achieved through a phenomenon called the enhanced permeability and retention (“EPR”) effect, which results from the typically leaky vasculature (increased permeation) and the impaired lymphatic drainage (increased retention) in the tumors. Consequently, NPs tend to remain intravascular in other tissues, yet they leak out of the vasculature in tumors and accumulate in the extra- vascular space.
  • EPR enhanced permeability and retention
  • the EPR effect is a molecular size-dependent phenomenon, where NPs with sizes above about 40 kDa usually display prolonged circulation due to slow renal clearance and possess the potential to gradually permeate tumors in a selective fashion.
  • the nanocarriers may encounter serum protein adsorption, leading to sequestration by the mononuclear phagocyte system ("MPS") and undesirable accumulation in the liver.
  • MPS mononuclear phagocyte system
  • Hyperbranched polyglycerol is a versatile nano-sized macromolecular platform that can be synthesized in a single step, resulting in polymers of controlled molecular weights with low polydispersity (Kainthan, R.K., et ah Macromolecules 2006; 39: 7708-7717; Imran ul-haq, M., et ah J. Polym. Set Part A: Polym. Chem. 2013; 51 : 2614-2621; Wilms, D., et ah Macromolecules 2009; 42: 3230-3236).
  • HPG Due to its structural similarities to polyethylene glycol (“PEG”), HPG possesses intrinsic ability to avoid MPS uptake, yet it is thermally and oxidatively more stable than PEG (Kainthan, R.K., et al. Biomaterials 2007; 28: 4581-4590). Furthermore, the hydroxyl groups of HPG can be derivatized with a variety of functional groups and hydrophobic molecules may be encapsulated in the hydrophobic core of an HPG NP (WO2006/130978).
  • PEG polyethylene glycol
  • HPG derivatized hyperbranched polyglycerol
  • NPs nanoparticles
  • HPG NPs disclosed herein may be used as an intravascular carrier for a
  • the HPG NPs disclosed herein may be used as an amphiphilic carrier for a chemotherapeutic agent for the treatment of cancer.
  • the claimed invention relates to a HPG nanoparticle for use in intravascular delivery of a biologically active moiety to a biological tissue
  • the HPG nanoparticle comprising a core comprising HPG derivatized with C 8 -C 18 alkyl chains and a shell comprising at least one hydrophilic substituent bound to hydroxyl groups of the core, wherein the at least one hydrophilic substituent comprises polyethylene glycol ("PEG"), methoxy polyethylene glycol (“MPEG”), or hyperbranched polyglycerol (HPG).
  • PEG polyethylene glycol
  • MPEG methoxy polyethylene glycol
  • HPG hyperbranched polyglycerol
  • the claimed invention relates to a method for intravascular delivery of a biologically active moiety to a biological tissue, the method comprising administering a HPG nanoparticle loaded with the biologically active moiety to a subject, wherein the HPG nanoparticle comprises a core comprising HPG derivatized with C 8 -C 18 alkyl chains and a shell comprising at least one hydrophilic substituent bound to hydroxyl groups of the core, wherein the at least one hydrophilic substituent comprises PEG or MPEG, or HPG.
  • the hydrophilic substituent bound to hydroxyl groups of the core is HPG
  • such HPG bound to hydroxyl groups of the core is not derivatized with alkyl chains.
  • the claimed invention relates to use of a HPG nanoparticle for intravascular delivery of a biologically active moiety to a biological tissue, the HPG
  • nanoparticle comprising a core comprising HPG derivatized with C -C 18 alkyl chains and a shell comprising at least one hydrophilic substituent bound to hydroxyl groups of the core, wherein the at least one hydrophilic substituent comprises PEG, MePEG, HPG.
  • the hydrophilic substituent bound to hydroxyl groups of the core is HPG
  • such HPG bound to hydroxyl groups of the core is not derivatized with alkyl chains.
  • use of such HPG NPs for manufacture of a medicament for such intravascular delivery of a biologically active moiety to a biological tissue.
  • the claimed invention relates to a pharmaceutical composition for use in intravascular delivery of a biologically active moiety to a biological tissue, the pharmaceutical composition comprising a HPG nanoparticle and a biologically active moiety, the HPG nanoparticle comprising a core comprising HPG derivatized with C 8 -C 18 alkyl chains and a shell comprising at least one hydrophilic substituent bound to hydroxyl groups of the core, wherein the at least one hydrophilic substituent comprises PEG, MPEG, or HPG.
  • the hydrophilic substituent bound to hydroxyl groups of the core is HPG, such HPG bound to hydroxyl groups of the core is not derivatized with alkyl chains.
  • the at least one hydrophilic substituent comprises PEG.
  • the C 8 -C 18 alkyl chains are C 8 and C 10 alkyl chains or C 10 alkyl chains.
  • the biologically active moiety is a hydrophobic drug.
  • the hydrophobic drug is a chemotherapeutic agent.
  • the chemotherapeutic agent is a taxane or an analog thereof.
  • the chemotherapeutic agent is docetaxel ("DTX") or an analog thereof.
  • the HPG nanoparticles may comprise about 0.05 to about 1 moles of the at least one hydrophilic substituent per mole of the HPG component of the core (i.e. the monomer unit glycidol in the polymer, and not including alkyl chains). In some embodiments, the HPG nanoparticles may comprise about 0.5 to about 0.6 moles of the at least one hydrophilic substituent per mole of the HPG component of the core. In some
  • the HPG nanoparticles may comprise 0.57 moles of the at least one hydrophilic substituent per mole of the HPG component of the core.
  • the HPG nanoparticles may comprise about 0.02 to about 1 moles of the C -C 18 alkyl chains per mole of the HPG component of the core. In some embodiments, the HPG nanoparticles may comprise about 0.4 to about 0.5 moles of the C 8 -C 18 alkyl chains per mole of the HPG component of the core. In some embodiments, the HPG nanoparticles may comprise 0.44 moles of the C 8 -C 18 alkyl chains per mole of the HPG component of the core.
  • the number average molecular weight of the hyperbranched polyglycerol nanoparticle may be about 10 kDA to about 1 M Da.
  • the number average molecular weight of the hyperbranched polyglycerol nanoparticle may be about 50 kDA to about 150 kDa.
  • the number average molecular weight of the hyperbranched polyglycerol nanoparticle may be about 70 kDA to about 128 kDa.
  • the polydispersity index of the hyperbranched polyglycerol nanoparticle may be about 1 to about 10.
  • the polydispersity index of the hyperbranched polyglycerol nanoparticle may be about 1 to about 1.5.
  • the composition of the HPG nanoparticles by weight is 19.8wt% HPG, 18.9 wt% Cio alkyl chains, and 61.3 wt% MPEG.
  • the biological tissue is a tumor.
  • HPG NPs disclosed herein may be described through a common nomenclature which identifies the basic hyperbranched structure, the core attributes, and the surface attributes as follows:
  • HPG-core(x)-shelli(yl)-shell 2 (y2)... -shell n (yn) (I) which designates a polymer consisting of HPG, comprising a core derivatized with a substituent selected from hydrophobic groups such as C 8 , C 10 , C 12 , C 14 , C 16 or C 18 alkyl groups that are either linear or branched or contain aryl substituents, wherein the amount of the core substituent is x, expressed in number of moles or as a percentage.
  • the polymer also has n substituents on the shell, such as HPG, PEG or MPEG, as disclosed herein.
  • Each shell substituent may be designated as being present in a certain amount yl, y2 oryn and can be expressed in number of moles or as a percentage.
  • general classes can be designated in the same manner, but without explicitly identifying the amounts of each.
  • the shell when the shell is PEG or MPEG, it may be further defined by the chain length of this polymeric component, for example, MPEG350, PEG200, etc. For the general class however, the molecular weight may be omitted.
  • polymers disclosed herein are meant to include all racemic mixtures and all individual structural isomers or variants, in particular as defined by the branch patterns within the HPG structure, or in terms of the physical attachment of the surface substituents to the HPG.
  • Figure 1 shows schematic diagrams of HPG-C 10 -HPG, HPG-C 10 -PEG and HPG-104;
  • Figure 2 shows in vitro toxicity assays with human dermal neonatal fibroblasts
  • HDFn human dermal microvascular endothelial cells
  • HDMEC human dermal microvascular endothelial cells
  • HUVEC human umbilical vein endothelial cells
  • Figure 4 shows histopathological analyses of mice tissues 34 days following intravenous administration of PBS (pH 7.4) or HPG-C 10 -PEG;
  • Figure 5 shows body weights of mice injected with PBS (pH 7.4) or HPG-Cio-PEG
  • Figure 6 shows docetaxel ("DTX") release from HPG-C 10 -PEG in PBS (pH 7.4) at
  • Figure 7 shows DTX release from HPG-d 0 -HPG in PBS (pH 7.4) at 37 °C;
  • Figure 8 shows in vitro toxicity assays with HDFn, pancreatic cancer cells (“PANC 1”), breast cancer cells (“MDA-MB-231”) and non-small cell lung cancer cells (“HCC 827”) incubated with various concentrations of DTX and DTX/HPG-C 10 -HPG for 72 h;
  • Figure 9 shows ratios of IC 50 between cancer cells to HDFn exposed to DTX or DTX/HPG-C 10 -HPG;
  • Figure 10 shows relative cell viability of colorectal cancer cells ("HT-29”), HCC 827 and pancreatic cancer cells (“BxPC-3") exposed to DTX or DTX/HPG-C 10 -PEG at different concentrations for 72 h;
  • Figure 11 shows relative cell viability of HUVEC and HDFn exposed to DTX or DTX/HPG-C 10 -PEG at different concentrations for 72 h;
  • Figure 12 shows proliferation (XTT) assays of HDFn and HT-29 exposed to DTX or
  • FIG. 13 shows scintillation count of 3 H-DTX taken up by HDFn and HT-29 cells
  • Figure 14 shows biodistribution of HPG-C 10 -HPG in three xenograft models with scintillation counts expressed as percent injected dose ("ID") per gram of tissue together with results of statistical analyses;
  • Figure 15 shows biodistribution of HPG-C 10 -PEG in mice with subcutaneous models of HT-29 and HCC 827;
  • Figure 16 shows tumor-to-blood ratios at 24 h and 72 h for HT-29 and HCC 827;
  • Figure 17 shows tumor-to-liver ratios at 24 h and 72 h for HT-29 and HCC 827;
  • Figure 18 shows tumor-to-spleen ratios at 24 h and 72 h for HT-29 and HCC 827
  • Figure 19 shows tumor-to-muscle ratios at 24 h and 72 h for HT-29 and HCC 827;
  • Figure 20 shows biodistribution of HPG-C 10 -HPG, HPG-C 10 -PEG and HPG-104 after 24 h or 72 h in NOD-SCID mice implanted with BxPC-3 (results of statistical analyses are provided below each graph);
  • Figure 21 shows tumor-to-normal tissue ratios for HPG-C 10 -HPG, HPG-C 10 -PEG and
  • HPG-104 (the line indicates the mean value and results of statistical analyses are provided below each graft);
  • Figure 22 shows relative tumor volume as a function of number of days after implant for mice with BxPC-3 tumors treated with either a low dose or high dose of DTX in a clinical formulation or DTX/HPG-C 10 -PEG;
  • Figure 23 shows a 1H NMR spectrum (D 2 0, 300 MHz) of HPG-C 10 -PEG.
  • Figure 24 shows a GPC-MALLS profile of HPG-C 10 -PEG in 0.1 N NaN0 3 .
  • NPs Polymer nanoparticles
  • HPG hyperbranched polyglycerol
  • HPG NPs Synthesis of HPG NPs has been previously described, including the production of amphiphilic copolymers and amphophilic block copolymers, including derivatization with various functional groups and/or the production of copolymers and block copolymers (such as the addition of alkyl groups through ester linkages and the addition of polyalkylene glycol groups).
  • HPG NPs disclosed herein may include HPG NPs derivatized with substituents having functional groups such that the HPG NPs may be used for
  • intravascular delivery of a biologically active moiety to a biological tissue of a subject.
  • intravascular delivery is used as it is normally understood to a person of ordinary skill in the art and often refers to administration of a biologically active moiety to a subject by entry into one or more blood vessels.
  • the HPG NPs disclosed herein have a "core", which includes an initiator (for example, trimethyloyl propane (TMP)) and HPG.
  • the HPG core may be derivatized with C 8 - C 18 alkyl chains.
  • the "HPG component” of the core is intended to refer only to the monomer unit glycidol in the polymer, and is not intended to include the alkyl chains with which the HPG is derivatized.
  • the "core” may be prepared by adding a glycerol epoxide (the hyperbranching component monomer) to an alkyl epoxide (which imparts the hydrophobic nature to the "core”).
  • the "core” may be enclosed in a "shell", wherein the shell comprises at least one hydrophilic substituent bound to hydroxyl groups of the core.
  • the HPG nanoparticles may comprise about 0.05 to about 1 moles of the at least one hydrophilic substituent per mole of the HPG component of the core. In some embodiments, the HPG nanoparticles may comprise about 0.5 to about 0.6 moles of the at least one hydrophilic substituent per mole of the HPG component of the core. In some embodiments, the HPG nanoparticles may comprise 0.57 moles of the at least one hydrophilic substituent per mole of the HPG component of the core.
  • HPG NPs may include C 8 -C 18 alkyl chains, or other similar alkyl chains.
  • alkyl is used as it is normally understood to a person of ordinary skill in the art and often refers to monovalent saturated aliphatic hydrocarbon groups having from one to 20 carbon atoms, unless otherwise defined.
  • the hydrocarbon may be either straight-chained or branched and may contain cycloaliphatic or aryl substituents.
  • Alkyl chains may be selected from one or more of C 8 -C 18 alkyl chains.
  • the alkyl chains may be selected from C 8 , Cg, C 10 , C 11 ⁇ C 12 , C 14 , C 16 or C 18 alkyl chains.
  • the alkyl chains may be selected from C 8 and/or C 10 alkyl chains.
  • the alkyl chain or chains may comprise about 0.02 to about 1 moles of the C 8 -C 18 alkyl chains per mole of the HPG component of the core.
  • the HPG nanoparticles may comprise about 0.4 to about 0.5 moles of the C 8 -C 18 alkyl chains per mole of the HPG component of the core..
  • the HPG nanoparticles may comprise 0.44 moles of the C 8 -C 18 alkyl chains per mole of the HPG component of the core.
  • the composition of the HPG nanoparticles by weight is 19.8wt% HPG, 18.9 wt% C 10 alkyl chains, and 61.3 wt% MPEG.
  • composition of the HPG nanoparticles may be determined by NMR spectroscopy.
  • alkyl chain or chains selected for the core and their amount may depend on the intended use for the HPG NP.
  • initiator is used as it is normally understood to a person of ordinary skill in the art and often refers to a small molecule comprising an alkyl component and more than one, and preferably more than two, hydroxyl groups. However, the initiator may have three or four or more hydroxyl groups.
  • An example of an initiator is trimethyloyl propane (TMP).
  • amphiphilic refers to any substance or portion thereof which is more soluble in a non-polar solvent than in a polar solvent. Hydrophobicity can be conferred by the inclusion of non-polar groups in a molecule, including, but not limited to, long chain saturated and unsaturated aliphatic hydrocarbon groups and such groups substituted by one or more aromatic, cycloaliphatic or heterocyclic group(s).
  • Hydrophilic refers to any substance or portion thereof which is more soluble in a polar solvent than in a non-polar solvent. Hydrophilic characteristics derive from the presence of polar or charged groups such as carbohydrates, phosphate, carboxylic, sulfato, amino, sulfhydryl, nitro, hydroxyl and other similar groups.
  • the hydrophilic portion may comprise MPEG, PEG or HPG.
  • polyethylene glycol or "PEG” is used as it is normally understood to a person of skill in the art and often refers to such compounds having a molecular weight between about 200 to about 20,000, depending on the number of ethylene oxide units in the polymer chain. Preferred molecular weights are from about 200 to about 400, about 200 to about 1000 and about 200 to about 2000, although molecular weights of about 2000 to about 8000 may also be used.
  • methoxypoly(ethylene oxide) or "MPEG” is used as it is normally understood to a person of skill in the art and often refers to such compounds having a molecular weight between about 350 to about 10,000, depending on the number of ethylene oxide units in the polymer chain. Preferred molecular weights are from about 350 to about 550, about 350 to about 750 and about 350 to about 2000, although molecular weights of about 2000 to about 5000 may also be used.
  • HPG NPs can be synthesized by the anionic ring opening multibranching polymerization (ROMBP) of glycidol alone (to form HPG- 104 which does not contain any hydrophobic core or hydrophilic shell) or glycidol with octadecyl glycidyl ether or glycidol with octadecyl glycidyl ether and MPEG.
  • ROMBP anionic ring opening multibranching polymerization
  • (HPG-C 10 -HPG (which has a hydrophobic core and an HPG shell) may be synthesized in two-steps in a single reaction pot by the anionic ring opening polymerization of a glycidol with octadecyl glycidyl ether ( to form the hydrophobic core) followed by the polymerization of glyciol alone (to form the HPG shell).
  • HPG-Qo-PEG may be synthesized by the anionic ring opening polymerization of glycidol with an epoxide-containing co-monomer by a two-step procedure.
  • a mixture of glycidol and octadecyl glycidyl ether (ODGE) is polymerized using trimethylol propane (TMP) as the initiator at 95 °C.
  • TMP trimethylol propane
  • polymerization is continued in the same reaction pot by the addition of ct-epoxy-co-methoxy polyethylene glycol (MPEG 40 o-epoxide) to afford HPG-C 10 -PEG NPs with a hydrophobic core and a hydrophilic PEG shell.
  • MPEG 40 o-epoxide ct-epoxy-co-methoxy polyethylene glycol
  • HPG NPs may have similar molecular weights (about 100 kDa) with very low polydispersity and similar size (hydrodynamic radius of approximately 5 nm).
  • HPG-C 10 -PEG is comprised of 22 mol% of the C 10 alkyl chains (hydrophobic moiety) and 28.5 mol% of PEG chains, as determined by 1H NMR spectroscopic analysis.
  • the embodiments of the HPG NPs disclosed herein include all possible stereochemical alternatives, including those illustrated or described herein.
  • the HPG NPs disclosed herein include isomers such as geometrical isomers having different branch patterns.
  • the HPG NPs synthesized by methods disclosed herein are random branching HPG NPs and will contain glycerol monomers that are fully reacted, e.g., linked in three directions, or partially reacted, being linked to another monomer in one or two directions.
  • the presence of each branching architecture may be confirmed by analytical techniques (e.g., 2D NMR HSQC experiments).
  • the HPG NPs disclosed herein may be in the solvent addition form.
  • the HPG NPs may be associated with a non-stoichiometric amount of a solvent, water and/or buffers, typically expressed as a weight or volume percent.
  • the solvent may be, for example, and without limitation, a pharmaceutically acceptable solvent or other biocompatible solvent including ethanol, DMSO, propylene glycol, glycerol, PEG200, PEG300, Transcutol or Solutol.
  • the HPG NPs disclosed herein may be administered to a subject.
  • subject is used as it is normally understood to a person of ordinary skill in the art and may include a human, non-human primate, rat, mouse, cow, horse, pig, sheep, goat, dog, cat, rabbit, etc.
  • the subject may be suspected of having or at risk for having a cancer or other disease that can be treated by intravascular delivery of a biologically active moiety to a biological tissue.
  • a cancer for example, lung, pancreatic, breast or colon cancers
  • an inflammatory disease as well as other indications may be desired targets of the HPG NPs disclosed herein for the parenteral delivery of a drug or other biologically active moiety.
  • compositions and HPG NPs according to embodiments disclosed herein may be administered in any of a variety of known parenteral routes.
  • the HPG NPs may be administered parenterally, i.e., by intra-articular, intravenous, subcutaneous, intramuscular, intraperitoneal, intracerebral, or
  • the biologically active moiety can be of any class of hydrophobic molecule which can be solubilized and held within the hydrophobic core of the HPG NP.
  • Compound solubility can be measured and defined as per the United States Pharmacopoeia / The National Formulary standards and guidelines.
  • small molecules, proteins, antibodies, peptides or pharmaceutically acceptable salts thereof may be held within the hydrophobic core.
  • the hydrophobic molecules may be molecularly targeted inhibitors, e.g. PI3K, CDK, PLK inhibitors.
  • the HPG NPs disclosed herein may be used for intravascular delivery of a chemotherapeutic agent to a tumor.
  • the chemotherapeutic agent may be a taxane.
  • the chemotherapeutic agent may be docetaxel ("DTX") or an analog thereof.
  • the chemotherapeutic agent may be paclitaxel or an analog thereof.
  • the biologically active moiety may be mitomycin or an analog thereof. Mitomycin may include all mitomycin analogs. Mitomycin and analogs thereof may include, for example, mitomycin A, mitomycin B, mitomycin C, mitomycin D, mitomycin F, mitomycin G, mitomycin H, mitomycin K and analogs thereof.
  • the biologically active moiety may be mitomycin C.
  • the biologically active moiety may be mitomycin F.
  • the biologically active moiety may be valrubicin.
  • the biologically active moiety may be vinblastine.
  • the biologically active moiety may be cisplatin.
  • the biologically active moiety may be methotrexate.
  • the biologically active moiety may be doxorubicin or an analog thereof.
  • the biologically active moiety may be epirubicin.
  • the biologically active moiety may be gemcitabine.
  • the biologically active moiety may be everolimus.
  • the biologically active moiety may be suramin.
  • the biologically active moiety may be a combination of moieties.
  • the combination of moieties may be methotrexate, vinblastine, and doxorubicin (M-VAC).
  • the combination of moieties may be M-VAC and cisplatin.
  • the combination of moieties may be two or more of valrubicin, cisplatin, paclitaxel and DTX.
  • the chemotherapeutic agent may hydrophobically bind with the core of the HPG NPs disclosed herein, allowing for the delivery of the chemotherapeutic agent to a tumor.
  • the chemotherapeutic agent may then be released.
  • the drug/HPG NP complex may be taken up by cancer cells and/or stromal cells in the tumor microenvironment and therapeutic agent may be then released.
  • the hydrophilic shell may act to shield the hydrophobic core and solubilize the HPG NPs in the blood of the subject.
  • the HPG NPs disclosed herein may be used as an intravascular formulation for the treatment of a cancer.
  • the HPG NPs described herein may be used to treat lung cancer, pancreatic cancer, colon cancer prostate cancer, ovarian cancer, skin cancer, breast cancer, head and neck cancer, liver cancer, stomach cancer, sarcomas, or brain cancer.
  • a solid tumor may be treated with a HPG NP disclosed herein.
  • HPG NPs for intravascular drug delivery systems should show low toxicity to non-cancerous cells, including primary human cells.
  • the toxicity of the HPG NPs disclosed herein was tested to two different types of endothelial cells, human umbilical vein endothelial cells (“HUVEC”) and human dermal microvascular endothelial cells (“HDMEC”).
  • the toxicity of the HPG NPs disclosed herein was also tested to fibroblasts (e.g., human dermal neonatal fibroblasts
  • HDFn HPG-Cio-HPG
  • HPG-C 10 -PEG Different concentrations of HPG-Cio-HPG and HPG-C 10 -PEG were incubated with the cell lines for 72 h and XTT assays were carried out to assess cell viability.
  • the results of the in vitro assay of HPG-C 10 -HPG to assess the toxicity of this HPG NP to HUVEC, HDMEC and HDFn are shown in Figure 2.
  • the IC 50 values are shown in Table 2. No ill effect on cell proliferation was observed at 1 mg/mL of HPG-Cio-HPG to all three cell types.
  • Toxicity of HPG-C 10 -HPG to HUVEC is evident at 5 mg/mL; however, the effect on proliferation of HDFn and HDMEC was not as significant at this concentration. Notably, the lower plateau was not reached with the concentrations of HPG-C 10 -HPG tested for HDFn, therefore, the IC50 value for this cell line could not be calculated.
  • HPG-C 10 -PEG the results are shown in Figure 3. Cytotoxicity was observed with HDFn and HUVEC at 0.5 mg/mL, while it was absent in HDMEC. At 1 mg/mL, cytotoxicity was apparent in all the primary cell lines tested.
  • HPG-C 10 -PEG The IC 50 values of HPG-C 10 -PEG were found to be 0.96, 1.2 and 0.56 mg/mL for HDFn, HDMEC and HUVEC, respectively.
  • HPG-C 10 -PEG exhibited much lower cytotoxicity compared to other polymeric NPs such as polyamidoamine (PAMAM) dendrimers; for example, the cytotoxicity observed for generation 4 PAMAM dendrimers ranged from 50-300 ⁇ g/mL (Zhang, X., et al. J. Control Release 2014; 174: 209-216).
  • PAMAM polyamidoamine
  • HPG NPs disclosed herein The toxicity of the HPG NPs disclosed herein was also tested on mice after intravascular administration. Normal C57/BL6 mice were intravenously injected with 100 mg/kg of HPG-Qo-PEG, and no detectable toxicity was evident from histopathological examination of organs ( Figure 4) and body weights (Figure 5). Hematological analysis of the whole blood samples collected from the mice also did not show any significant difference in the major hematological parameters between the control and test groups (Table 3). The hematological evaluation is critical for safety since intravascular delivery of hydrophobic biologically active moieties is generally designed to be long-circulating and hence have long blood residence times. These results indicate that the HPG NPs disclosed herein can be safely injected intravenously into mice without any adverse effects and may be blood compatible. Thus, the HPG NPs disclosed herein may be intravascularly injected into a subject without any or minimal adverse effects.
  • the HPG NPs disclosed herein may be loaded with a biologically active moiety.
  • the biologically active moiety may interact with the hydrophobic core of the HPG NP and be encapsulated within the hydrophilic shell of the HPG NP.
  • the content and type of alkyl chains at the core may affect the solubility of the HPG NPs and the ability of HPG NPs to carry a hydrophobic drug. The higher the content of alkyl chains, the more likely it is that a hydrophobic drug will be carried in the core of the HPG NP. However, if the content of alkyl chains is too high, the HPG NP will no longer be soluble in the blood stream.
  • the content of alkyl chains in the HPG NP is about 0.02 to about 1 moles of alkyl chains per mole of the HPG component of the core.
  • the HPG nanoparticles may comprise about 0.4 to about 0.5 moles of alkyl chains per mole of the HPG component of the core.. In some embodiments, the HPG nanoparticles may comprise 0.44 moles of alkyl chains per mole of the HPG component of the core.
  • HPG NPs disclosed herein and loaded with a chemotherapeutic agent may show a prolonged drug release profile.
  • drug loading experiments were undertaken to load up to 5 wt % of 3 H-DTX into HPG-C 10 -PEG, with the highest stability of the formulation achieved at 1% loading, which translates to a DTX:HPG-C 10 -PEG ratio of 1 : 1.
  • An in vitro drug release profile in PBS (pH 7.4) at 37 °C from HPG-C 10 -PEG is shown in Figure 6.
  • the release profile of 3 H-DTX suggests that the drug was released in a sustained and controlled manner over about 17 days, with 50% of the drug released in about 3 days and 90% of the drug released within about 15 days.
  • HPG NPs disclosed herein and loaded with a chemotherapeutic agent such as DTX may show a relative protective effect with non-cancerous cells, potentially widening the therapeutic index of the HPG NPs disclosed herein.
  • the HPG NPs disclosed herein which are loaded with a chemotherapeutic agent may effectively kill cancer cells while showing less effect on normal cells.
  • HPG NPs disclosed herein and loaded with a chemotherapeutic agent may show a preferential protective effect on primary noncancerous cells.
  • HPG NPs comprising HPG-C10-PEG appears similar for both primary cells and cancer cells, indicating that the enhanced killing of cancer cells compared to primary cells may be related to differences in how HPG NPs are processed intracellularly.
  • the cellular uptake rates of HPG-C10-HPG by normal and cancer cells differ, which suggests that cellular uptake also depends on the structure and composition of the HPG NPs.
  • the ability of the HPG NPs disclosed herein and loaded with DTX to kill cancer cells was assessed with XTT assays.
  • the ratio When the ratio is less than 1, the drug is more toxic to normal cells than to cancer cells.
  • the ratios can be plotted graphically and tabulated, as shown in Figure 9 and Table 4.
  • the IC 50 value for HDFn was lower than those for HCC 827 and PANC 1, indicating that DTX can be more toxic to normal fibroblasts than to cancer cells.
  • MDA-MB-231 cells were more susceptible to DTX than HDFn.
  • the IC 50 ratios of DTX for the three cancer cell lines tested were very close to 1.
  • the IC 50 ratios for HCC 827, PANC 1 and MDA-MB-231 cells were 2.1, 3.7 and 3.3, respectively.
  • DTX/HPG-C 10 -PEG For experiments conducted with DTX/HPG-C 10 -PEG, DTX, a drug that targets microtubules, displayed similar toxicity to all cells tested ( Figures 10 and 1 1), as expected.
  • DTX/HPG-C 10 -PEG killed cancer cells (colorectal cancer cells ("HT-29”), HCC 827 and pancreatic cancer cells (“BxPC-3”)) with similar effect as the free DTX ( Figure 10) while the primary human cells (HUVEC and HDFn) were not as susceptible to killing by DTX/HPG- Cio-PEG ( Figure 1 1).
  • the HPG NPs disclosed herein may show a protective effect on normal cells compared to free DTX.
  • HPG NPs disclosed herein may enhance the therapeutic ratio for the chemotherapeutic drug by killing the cancer cells while protecting the normal cells from cytotoxic effects of DTX.
  • the differential killing effects of DTX/HPG-Cio-PEG suggest two non-exclusive possibilities. Firstly, the uptake of DTX/HPG-Qo-PEG differs between the cancer cells and the primary human cells thereby leading to a difference in toxicity effects. Secondly, there is a difference between the processing of DTX/HPG-C ⁇ -PEG in the cancer cells, when compared to the non-cancerous cells, so that the availability of DTX is different. To understand if the uptake mechanism of DTX/HPG-C 10 -PEG is energy-dependent, cellular uptake experiments were performed at different temperatures.
  • NPs for intravascular drug delivery have been their uptake by the mononuclear phagocyte system ("MPS") as a result of rapid sequestration of NPs from the blood by hepatic midzonal and periportal Kupffer cells
  • Typical NPs undergo high uptake into the organs of the MPS such as the liver and spleen.
  • Previous studies with PEGylated NPs of similar size have shown mean liver and spleen uptake of
  • HPG NPs disclosed herein may have a more favourable biodistribution profile due to reduced MPS clearance and long circulation half-life. In various embodiments, the HPG NPs disclosed herein may persist in the blood circulation of a subject and may also show low accumulation in the liver and spleen of the subject.
  • HPG NPs disclosed herein may accumulate in a tumor. However, if the hydrophobic core is not sufficiently shielded by the hydrophilic shell, HPG NPs may accumulate in the liver and spleen. Without being bound by any particular theory, it appears that without significant shielding, the hydrophobic core of the HPG NPs disclosed herein interacts with cells of the MPS.
  • FIG 14 shows blood and tumor distributions of radio-labelled HPG-C 10 -HPG in three subcutaneous xenograft models (HCC 827, HT-29 and BxPC-3). Results for a similar study with HPG-Cio-PEG are shown in Figure 15. These xenograft models were chosen to reflect the heterogeneity of the tumor microenvironments.
  • BxPC-3 is a xenograft model with a low degree of vascularization and vascular permeability.
  • HT-29 has high vascularity and increased vascular permeability.
  • HCC 827 is a non-small cell lung carcinoma that is known to recruit fibroblasts.
  • HPG-C 10 -PEG For HPG-C 10 -PEG, similar patterns of biodistribution were observed for two xenograft models (HT-29 and HCC 827) ( Figure 15).
  • the amount of HPG-C 10 -PEG in the circulation decreased significantly from 24 h to 72 h post-injection, while the accumulation of HPG-C 10 - PEG in the tumors was sustained.
  • the mean liver and spleen uptake for HPG-C 10 -PEG in the xenograft models ranged between 9-16% and 4-7 % ID/g, respectively, while the mean tumor uptake of HPG-C 10 -PEG was in the range of 3-6 % ID/g at the 24 h and 72 h time points.
  • Tumor-to-blood ratios indicated increasing accumulation in the tumors over time (Figure 16).
  • the significant tumor uptake was likely attributable to the ability of the HPG-Cio-PEG to circulate in the blood for a long duration as well as to the small hydrodynamic diameter of approximately 12 nm of HPG-C 10 -PEG.
  • Tumor-to-liver and tumor-to-spleen ratios of HPG- Cjo-PEG ( Figures 17 and 18, respectively) also suggest relatively low elimination by the MPS on the basis that there was minimal difference between the two xenograft models.
  • Long circulation half-life of up to 34 h has previously been observed for a similar class of NPs (Kainthan, R. K., et al. Biomaterials 2008; 29: 1693-1704).
  • Very little HPG-C 10 -PEG accumulated in skeletal muscles, which is evident with the large tumor-to-muscle ratios in both models (Figure 19).
  • HPG-C 10 -PEG may act as a drug carrier in a similar way, while being easier to produce and a more versatile carrier for intravascular drug delivery.
  • HPG- o- PEG may allow for bulk production at an acceptable cost while also addressing the safety concerns over prion and viral transmission in the blood supply associated with animal-derived albumin.
  • biodistribution profile of the HPG NPs disclosed herein may be achieved through the enhanced permeability and retention
  • FIG. 20 shows the results of biodistribution at 24 and 72 h post-injection for HPG-Cio-HPG, HPG-C 10 -PEG and HPG-104.
  • the biodistribution studies were performed with the same tumor xenograft model (BxPC-3 subcutaneously implanted into NOD-SCID mice). Similar to the biodistribution with HT-29 and HCC 827 xenograft models, HPG-Cio-HPG accumulated heavily at the liver and the spleen.
  • HPG- C 10 -PEG the accumulation of HPG- C 10 -PEG at the liver and the spleen was significantly reduced and the proportion of blood- resident HPG-C 10 -PEG at 24 and 72 h was increased significantly.
  • HPG- 104 showed the lowest accumulation at the liver and the spleen.
  • the results suggest that the unfavourable accumulation of HPG-C 10 -HPG at the liver and spleen may be due to the presence of the hydrophobic core, whose effect is partially shielded by PEGylation (HPG-C 10 -PEG) and its removal (HPG-104) results in a significant decrease of accumulation at these organs.
  • the HPG NPs disclosed herein may deliver chemotherapeutic agents to tumors while sparing the normal tissue (such as the liver) to reduce or minimize nonspecific toxicity.
  • HPG-C 10 -HPG, HPG-C 10 -PEG and HPG-104 may all accumulate in tumors at similar levels at 24 and 72 h (e.g., 3-6% ID/g).
  • the ratio of % ID/g of the HPG NPs between the liver and the tumor for each mouse implanted with an HPG NP may be a useful indicator of specific accumulation.
  • Figure 21 shows the tumor-to-liver, tumor-to-spleen, tumor-to-heart and tumor-to-muscle ratios of HPG-C 10 -HPG, HPG-C 10 -PEG and HPG-104 for assessing specific tumor accumulation.
  • the tumor-to-liver and tumor-to-spleen ratios of HPG-C 10 -HPG was the lowest among the three HPG NPs; however, the tumor-to-heart ratio of HPG-C 10 -HPG was the highest with good tumor-to-muscle ratio.
  • chemotherapeutic agent may deliver the chemotherapeutic agent to a tumor and decrease a volume of that tumor.
  • a low dose (5 mg/kg) (“LD") of DTX delivered intravenously as DTX in a clinical formulation (sourced from Hospira) or HPG-C 10 -PEG loaded with comparable amount of DTX (DTX/HPG-Cio-PEG) showed a decreased relative tumor volume compared to those mice treated with PBS (pH 7.4).
  • tumor volume for mice treated with DTX/HPG-Cio- PEG was significantly lower than for mice treated with the clinical formulation of DTX at the low dose.
  • HD high doses of DTX (15 mg/kg)
  • three out of five mice treated with the clinical formulation of DTX lost greater than 20% of their body weight and needed to be euthanized about 60 days following implantation of the BxPC-3 cells.
  • HDs of DTX incorporated into HPG-Cio-PEG could be delivered to a tumor and effectively decreased the relative tumor volume, indicating that DTX-HPG-C 10 -PEG may be more efficacious than clinical formulations of DTX.
  • Suitable pharmaceutical compositions comprising the HPG NPs disclosed herein may be formulated by means known in the art and their mode of intravascular administration and dose determined by the skilled practitioner. Many techniques known to one of skill in the art are described m Remington: the Science and Practice of Pharmacy by Alfonso Gennaro, 20 ed., Lippencott Williams & Wilkins (2000).
  • an “effective amount” of a pharmaceutical composition disclosed herein includes a therapeutically effective amount or a prophylactically effective amount.
  • a “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result, such as decreased cancer cell proliferation, increased life span or increased life expectancy.
  • a therapeutically effect amount of a HPG NP incorporating a biologically active agent may vary according to factors such as the disease state, age, sex, and weight of the subject, and the ability of the biologically active agent to elicit a desired response in the subject. Dosage regimens may be adjusted to provide the optimum therapeutic response.
  • a therapeutically effective amount is also one in which any toxic or detrimental effects of the formulation are outweighed by the therapeutically beneficial effects.
  • prophylactically effective amount refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result, such as the prevention or the prevention of the progression of an indication.
  • a prophylactic dose is used in subjects prior to or at an earlier stage of disease.
  • dosage values may vary with the severity of the condition to be alleviated.
  • specific dosage regimens may be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions.
  • the amount of composition may vary according to factors such as the disease state, age, sex and weight of the subject. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It may be advantageous to formulate compositions in dosage unit form for ease of administration and uniformity of dosage.
  • HPG NPs disclosed herein may be used, for example, and without limitation, in combination with other treatment methods.
  • HPG NPs disclosed herein incorporating a chemotherapeutic agent may be used as neoadjuvant (prior), adjunctive (during), and/or adjuvant (after) therapy in combination with other therapies known to one of ordinary skill in the art, including additional chemotherapies, radiotherapy, and/or surgery.
  • Octyl/decyl glycidyl ether (ODGE) was purified by distillation under reduced pressure, a- Epoxy-co-methoxy polyethylene glycol was synthesized as per published procedure (Cho, K., et al. Macromol. Rapid Commun. 1999, 20: 598-601). Tritiated methyl iodide was purchased from ARC Radiochemical (St. Louis, MO) as solution in toluene and was used directly after dilution in dimethylsulfoxide.
  • GPC Chromatography
  • MALLS multi- angle laser light scattering
  • Optilab DSP refractive index detector both from Wyatt Technology.
  • Hydrodynamic radii (R h ) of the polymers were measured using quasi-elastic light scattering (QELS) detector coupled to the MALLS detector.
  • QELS quasi-elastic light scattering
  • GPC analysis was performed using Waters ultrahydrogel columns (guard, linear and 120) and 0.1 N NaN0 3 (10 mM phosphate buffer) as the mobile phase. All animals were bred in the Animal Resources Centre at the British Columbia Cancer Research Centre and all experimental procedures were done in accordance with ethical guidelines set by the Animal Care Committee and the Canadian Council on Animal Care.
  • a-Epoxy-co-methoxy polyethylene glycol (mPEG 4 oo-epoxide, 54 mL) was added slowly over a period of 24 h. After complete addition of the monomer, the reaction mixture was stirred for an additional 3 h. The polymer was dissolved in methanol and the solution was passed through Amberlite-IER-120 resin to remove the potassium ions. The unreacted ODGE was removed by extraction with hexane. The polymer was precipitated from excess diethyl ether, dissolved in water and dialyzed against water using MWCO 10000 cellulose acetate membrane for 2 days with periodic changes in water. The dry polymer was recovered by freeze drying.
  • the polymer was synthesized using a similar procedure as that described above with only glycidol as the monomer.
  • the polymer does not contain C 10 alkyl groups or PEG.
  • GPC-MALLS (0.1 N NaN0 3 ): M erase: 104000, M w /M n : 1.15, R h : 5.1 nm.
  • DTX loading into HPG-C 10 -HPG and HPG-C 10 -PEG was carried out as described previously (Mugabe, C, et al. Int. J. Pharm. 2011 ; 404: 238-249). Briefly, an appropriate amount of HPG-C 10 -HPG or HPG-C 10 -PEG was weighed in a 4 mL vial and dissolved in 1 mL of methanol or acetonitrile, respectively. To determine optimal drug loading, as part of separate experiments, DTX at up to 5 % w/w (of HPG-C 10 -HPG or HPG-C 10 -PEG) was added to the vial.
  • HDFn, HUVEC and HDMEC cells were purchased from Life Technologies (Carlsbad, CA). Only early passages of the cells were used in experiments. All cancer cells (BxPC-3, HT-29, HCC 827, PANC1 and MDA-MB-231) were purchased from American Type Cell Culture (ATCC). All cells were cultured according to the recommendation of the suppliers and tested regularly for mycosplama contamination (InvivoGen, San Diego, CA).
  • mice Female C57/B6 mice were intravenously injected with 100 mg/kg of HPG-C 10 -PEG or PBS (pH 7.4) at an injection volume of 100 ⁇ 1/20 g through the tail vein. Body weights of mice and signs of ill health were monitored for more than 30 days. There were no behavioural or pathological signs in the animals throughout the study period. The mice were then euthanized and all the major organs such as brain, lungs, heart, stomach, small intestine, ovaries, kidneys, spleen, urinary bladder, liver and skeletal muscle were harvested. The organs were fixed with 10 % formalin, embedded in paraffin, sectioned, stained with hematoxylin and eosin, and evaluated by a board-certified pathologist.
  • HPG-C 10 -HPG and HPG-C 10 -PEG were each loaded with DTX (as described above) spiked with tritium-radiolabeled DTX ( H-DTX, Moravek Biochemicals, Brea, CA).
  • the formulation (1 mL) was placed into biodialyser units (Sigma- Aldrich, Oakville, Ontario) comprising 10,000 Da cut off membranes.
  • the biodialyser units were placed in 500 mL PBS (pH 7.4) at 37 °C under continuous stirring and drug release was measured at pre-determined time points.
  • HDFn and HT-29 cells were seeded in 96-well plates the day before experimentation.
  • Cells were pre-incubated on ice (4 °C) or at 37 °C for 3 h, after which the culture media was removed and replaced with media containing DMSO, dilutions of DTX or DTX-loaded HPG- Cto-PEG. Cells were further incubated on ice or at 37 °C for 3 h before the media was removed and washed once with media. Six replicates were included for each test condition. One hundred microliters of fresh media were dispensed into each well and the cells were incubated for 72 h before XTT assays were carried out. Four independent experiments were carried out, and significant difference was determined with paired (with the same experiment) t-test. Error bars in Figure 12 are standard errors of the mean.
  • HDFn and HT-29 cells were seeded in 12-well plates at 50,000 or 100,000 cells/well. On the next day, the media was removed and the media containing H-DTX or ⁇ -DTX-loaded HPG-C 10 -PEG was added to the wells. Triplicates were performed for each test condition and three independent experiments were carried out. After 3 h of incubation at 37 °C, the cells were washed with PBS (pH 7.4) and solubilised with NaOH. The lysate samples were mixed with scintillation cocktail and scintillation counts were taken as readout of 3 H-DTX taken up by the cells.
  • Percent uptake was computed by scintillation counts of cell lysate normalized to scintillation counts of amount of 3 H-DTX or 3 H-DTX-loaded HPG- C 10 -PEG applied to the wells. Error bars in Figure 13 are standard errors of the mean.
  • Radiolabeling of HPG-C 10 -PEG was performed by partial conversion of the hydroxyl groups to methoxide groups using tritiated methyl iodide (C 3 H 3 I).
  • C 3 H 3 I tritiated methyl iodide
  • Two hundred milligrams of HPG-C 10 -PEG was dissolved in 2 mL of l-methyl-2-pyrrolidinone (NMP) and approximately 5% of the hydroxyl groups were deprotonated using sodium hydride.
  • NMP l-methyl-2-pyrrolidinone
  • a calculated amount of tritiated methyl iodide (toluene solution) dissolved in DMSO was added to this solution so as to achieve methylation of approximately 1% of the hydroxyl groups.
  • the reaction mixture was stirred at room temperature for 15 h, 15 mL of water was added and the labelled HPG NP was purified by dialysis against water using MWCO 1000 dialysis membrane until the dialysate contained low amounts of radioactivity, this took approximately 48 h.
  • the HPG NP solution was then filtered through 0.2 ⁇ syringe filter and the HPG NP weight was determined from the total volume and the HPG NP concentration from a known volume of the solution after freeze-drying.
  • the HPG NP solution for the animal studies was prepared by the addition of appropriate amounts of NaCl and water to achieve the desired osmolarity and the specific activity was measured by scintillation counting.
  • mice Male non-obese diabetic-severe combined inrmunodeficient (“NOD-SCID”) mice of 8- 12 weeks of age were implanted subcutaneously with 5 x 10 6 cancer cells per mouse and monitored. When the tumours reached the size of about 200-400 mm 3 , tritium-labelled HPG- Cio-HPG, HPG-Cio-PEG or HPG-104 was injected intravenously through the tail vein of each mouse (200 ⁇ 1/20 g mouse, 20 mg/kg). At 6, 24 and 72 h post-injection, the mice were euthanized with blood, tumors and major organs removed for processing and scintillation counts. Three subcutaneous xenograft models were employed (HCC 827, BxPC-3 and HT-29).
  • the samples were allowed to cool before the addition of 50 ⁇ , of 200 mM EDTA, 200 uL 30% H 2 0 2> and 25 ⁇ 10 N HC1 to each sample. The samples were then incubated at room temperature for 1 h prior to the addition of 5 mL of scintillation cocktail per sample. Samples were analyzed by scintillation counting. For reference, 20 ⁇ of the tritiated HPG stock in triplicate was analyzed by scintillation counts for specific activity determination. All samples were counted at least 1 week after the processing. Distribution of the tritiated HPG in tissues are reported as percent injected dose per gram of tissue (% ID/g).
  • NOD-SCID mice of 8-12 weeks of age were implanted subcutaneously with 5 x 10 6 BxPC-3 cells per mouse and monitored. After 44 days, a low dose (5 mg/kg) or high dose (15 mg/kg) of the clinical formulation of DTX (again, sourced from Hospira) or DTX/HPG-C 10 - PEG was injected intravenously through the tail vein of each mouse.
  • DTX sourced from Hospira
  • DTX/HPG-C 10 - PEG was injected intravenously through the tail vein of each mouse.
  • follow-up doses were also administered 51 days and 58 days following implantation of the cancer cells.
  • a control group was used which received PBS (pH 7.4). The relative tumor volume was monitored until approximately 75 days following implantation of the cancer cells.
  • mice were treated with phosphate-buffered saline only (sham treatment);
  • mice were treated with Taxotere at 0.5 mg/mL, 200 uL/20 g mouse weight;
  • mice were treated with Taxotere at 1.5 mg/mL, 200 uL/20 g mouse weight;
  • mice were treated with HPG-C 10 -PEG loaded with 0.5 mg/mL of Docetaxel, 200 uL/20 g mouse weight; and
  • mice were treated with HPG-C 10 -PEG loaded with 1.5 mg/mL of Docetaxel, 200 uL/20 g mouse weight.

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Abstract

L'invention concerne des polyglycérols hyper-ramifiés (HPG) dérivés destinés à être utilisés en tant qu'agents pour l'administration d'un médicament ou autre fragment biologiquement actif à un tissu cible pour traiter des indications telles que le cancer, qui peuvent être utiles dans le traitement ou la fabrication d'un médicament, dans la préparation d'une composition pharmaceutique pour le traitement d'indications comme le cancer. L'invention concerne également des HPG dérivés, pour une utilisation en tant que pré-traitement ou co-traitement pour améliorer l'absorption d'un médicament dans un tissu. En outre, l'invention concerne des procédés de fabrication de HPG dérivés. Dans des modes de réalisation particuliers, les HPG sont dérivés avec des chaînes d'alkyle en C10. Ces HPG dérivés peuvent être appropriés pour l'administration par voie intravasculaire.
PCT/CA2015/050600 2014-06-30 2015-06-26 Polyglycérol hyperramifié à dérivatisation hydrophobe pour l'administration d'un médicament par voie intravasculaire Ceased WO2016000070A1 (fr)

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CN109021229A (zh) * 2018-08-06 2018-12-18 武汉大学 一种含硫超支化聚缩水甘油醚共聚物的制备方法及其应用
CN109021229B (zh) * 2018-08-06 2020-01-24 武汉大学 一种含硫超支化聚缩水甘油醚共聚物的制备方法及其应用
CN109608647A (zh) * 2018-12-25 2019-04-12 上海交通大学医学院 活性氧响应的聚合物、载体及其应用
CN109608647B (zh) * 2018-12-25 2022-01-04 上海交通大学医学院 活性氧响应的聚合物、载体及其应用
CN113444250A (zh) * 2021-06-18 2021-09-28 绍兴文理学院附属医院 一种含有聚谷氨酸基团的聚甘油脂肪酸酯衍生物及其合成方法和其在药物制剂中的应用
CN114848834A (zh) * 2022-05-25 2022-08-05 浙江大学医学院附属第一医院 一种双药物共递的复合多层纳米载体及其制备方法和应用
CN114848834B (zh) * 2022-05-25 2024-01-26 浙江大学医学院附属第一医院 一种双药物共递的复合多层纳米载体及其制备方法和应用

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