Release Behavior of 5-Fluorouracil from Chitosan-Gel Nanospheres Immobilizing 5-Fluorouracil Coated with Polysaccharides and Their Cell Specific Cytotoxicity

Abstract

Small-sized chitosan-gel nanospheres, CNSs (average diameter 250 nm), containing 5-fluorouracil (5FU) or immobilizing 5FU derivatives (aminopentyl-carbamoyl-5FU or aminopentyl-ester-methylene-5FU) were prepared by the glutaraldehyde crosslinking technique and the emulsion method. When chitosan was crosslinked with glutaraldehyde, these 5FU derivatives were simultaneously immobilized to CNSs by means of Schiff's base formation. The CNSs were coated with anionic polysaccharides, such as 6-O-carboxymethyl-N-acetyl-α-l,4-polygalactosamine/Na (CM-NAPGA/Na), 6-O-carboxymethyl-chitin/Na (CM-chitin/Na), and sodium hyaluronate, through formation of a polyelectrolyte complex membrane to give CNS/polyanion, i.e., CNS/G, CNS/C, and CNS/H, respectively. The polyelectrolyte complex of polysaccharide was employed to achieve the controlled release and effective targeting of 5FU by the CNSs. The release rate of 5FU from the CNSs could be controlled by immobilization of 5FU, degree of deacetylation of chitosan used and coating with polysaccharides. Since very few galactosamine residues are known to be able to cross-react with ligands for galactose, the galactosamine residues on the surface of CNS/Gs are expected to act as the targeting moieties for hepatocyte. The CNS/G showed the lectinmediated aggregation phenomenon by the addition of APA lectin. Moreover, CNS/G had the highest cytotoxic activity among the three kinds of CNS/polyanion and CNS in HLE human hepatoma cell culture system in vitro.     

Antitumor Activity of Vinyl Polymer Immobilized 5-Fluorouracils through Organosilicon-Amine Spacer Groups via Urea Bonds

Abstract

The homopolymerization of vinyl monomer immobilized 5-fluorouracils (5FU) through organosilicon-amine spacer groups via urea bonds (KY-20) and the copolymerization of KY-20 with vinyl monomer were carried out by a radical technique to give the homopolymer [Poly(KY-20)] and copolymer [Poly(KY-20/vinyl monomer)], respectively. The antitumor activities of Poly(KY-20) and Poly(KY-20/vinyl monomer) were tested against p388 lymphocytic leukemia by intraperitoneal (i.p.) transplantation/i.p. or oral (p.o.) administration and against Meth-A fibrosarcoma or MM46 mammary adenocarcinoma in mice by subcutaneous (s.c.) implantation/p.o. administration. These 5FU/ organosilicon-amine hybrid polymer conjugates exhibited significant survival effects against p388 leukemia in mice i.p./i.p. or i.p./p.o. Poly(KY-20/HPMA) also showed stronger growth-inhibitory effects against Meth-A fibrosarcoma and MM46 mammary adenocarcinoma in mice s.c./p.o.     

Design of Chitosan-5Fu Conjugate Exhibiting Antitumor Activity

In order to provide a macromolecular prodrug of 5-fluorouracil (5FU) reducing the side effects, having an affinity for tumor cells, and exhibiting strong antitumor activity, the covalent attachments of 5FUs to chitosan and chitosamino-oligosaccharide (COS) through hexamethylene spacer groups via urea, urea bonds were carried out. The effect of prolongation of life was tested in vivo against p388 lyrnphocytic leukemia in female CDF, mice by intra-peritoneal (i.p.) transplantation/i.p. injection and the growth-inhibitory effect on Meth-A fibrosarcoma or MH-134Y heputoma was evaluated in vivo in SPF-C3H/He scl male or Balb/c male mice by subcutaneous (s.c.) implantation/intravenous (i.v.) injection. The effects of the degree of polymerization of chitosan and     

Adsorption of polyfunctional 5‐fluorouracil and 2,4‐dithio‐5‐fluorouracil on Au(111) surface: Structure,energy, and electronic transmission

Adsorption of 5‐fluorouracil (5‐FU) and 2,4‐dithio‐5‐fluorouracil (2,4‐DT‐5‐FU) on Au(111) surface at low coverage is studied by using periodic‐slab‐density functional theory calculation. Isolated 5‐FU molecule adsorbs preferentially at bridge site in a vertical configuration via N? H group by forming the N? H···Au nonconventional H‐bond. The formation of the anchor Au? O bond is not observed. Substitution of oxygen atoms of 5‐FU with sulfur strongly influences the nature of adsorption and leads to the Au? S anchor bond and the N? H···Au nonconventional H‐bond of single 2,4‐DT‐5‐FU molecule on Au(111) surface. The adsorption site and orientation of 2,4‐DT‐5‐FU molecule on the surface are similar to those of 5‐FU. The metal–molecule coupling effects at asymmetric Au/S(N? H)S/mol/C? H/Au and Au/N? H/mol/O/Au transport junctions and symmetric Au/S(N? H)S/mol/mol/S(N? H)S/Au and Au/O/mol/mol/O/Au transport junctions are also investigated. The electronic structure is analyzed in detail, and the obtained results are used for illustrating the electron transmission in metal–molecule–metal systems. © 2011 Wiley Periodicals, Inc. Int J Quantum Chem, 2011     

Preparation and characterization of microwave-treated carboxymethyl chitin and carboxymethyl chitosan films for potential use in wound care application

CM-chitin and CM-chitosan films were successfully crosslinked by microwave treatment. Crosslinking of the microwave-treated CM-chitin films involved mainly the carboxylate and the secondary alcohol groups, while crosslinking of microwave-treated CM-chitosan films involved the carboxylate and the amino groups. In addition, the crystallinity of CM-chitin increased with increasing microwave treatment time, whereas an increase in the crystallinity of the microwave-treated CM-chitosan films was not observed. At a similar percentage of weight loss, the crosslinking of either CM-chitin or CM-chitosan films by microwave treatment required much less stringent condition when compared with the crosslinking by autoclave treatment. Based on both direct and indirect cytotoxicity assays, the cytotoxicity of the microwave-treated CM-chitin films was negative, while that of the microwave-treated CM-chitosan films was positive. Human fibroblasts adhered on the surface of microwave-treated CM-chitosan films much better than on the surface of microwave-treated CM-chitin films.     

6-O-carboxymethyl-chitin (CM-chitin) as a drug carrier

Gel was prepared from 6-O-carboxymethyl-chitin (CM-chitin) by the addition of iron(III) chloride under mild conditions without any organic solvent. The optimal conditions for the gel formation were 15 to 30 mM iron(III) chloride and 0.5 to 0.8 degree of substitution in CM-chitin. The amounts of bovine serum albumin (BSA) and the anticancer drug doxorubicin (DOX) incorporated into CM-chitin gels were more than 80% and 30%, respectively under the conditions described above. The release of BSA or DOX from the gels was observed to be increased by lysozyme digestion in a time-dependent manner. This result indicates that CM-chitin might prove useful as a carrier gel for the sustained release of drugs and cytokines, including vaccines.     

Syntheses of 5-Fluorouracil-Terminated Monomethqxypqly(Ethylene Glycol)S,Their Hydrolysis Behavior,and Their Antitumor Activities

The covalent attachments of 5-fluorouracil (5FU) units to poly(ethylene glycol) monomethoxy ether (MeO-PEG) attached through ester, amide, and ether bonds were carried out; three types of linkages were obtained through which 5FU units were attached to MeO-PEG. For the investigation of the release of the 5FU units, the homogeneous hydrolysis was investigated in vitro in the presence and absence of enzymes. Although the rate of release of l-β-carboxyethyl-5FU or 1-β-hydroxyethyl-5FU from the compounds was fast, the release of 5FU itself was slow. In addition, the antitumor activity of these three types of MeO-PEG-bound 5FU unit was tested in vivo by preliminary screening by the National Cancer Institute or by the Japanese Foundation for Cancer Research.     

Dendritic polyaniline nanoparticles synthesized by carboxymethyl chitin templating

Dendritic polyaniline (PANI) nanoparticles were synthesized via oxidative polymerization of aniline, using ammoniumperoxodisulfate as an oxidant, and CM-chitin as a template. The reaction was performed under acidic conditions and the template was removed after the polymerization was completed. Molecular characterization (including UV-vis, FTIR, TGA, and XRD) suggests that the structure of the synthesized dendritic PANI nanoparticles is identical to that of the emeraldine form of PANI, synthesized by the conventional route (without the addition of the CM-chitin template). SEM images reveal that the dendritic PANI nanoparticles have an average diameter in the nanometer range, and are globular in shape, with radially oriented PANI dendrites; in contrast, irregularly-shaped aggregates of PANI are obtained using the conventional synthesis. It was further found that the size of the dendritic PANI nanoparticles is dependent on the CM-chitin content. The higher the CM-chitin concentration, the smaller is the size of the dendritic PANI nanoparticles obtained. An interpretation of these observations and a possible formation mechanism are proposed based on self-assembly between the CM-chitin chains and the aniline monomer.     

Physical and Electrical Properties of Chlorophyllin/Carboxymethyl Chitin and Chlorophyllin/Carboxymethyl Chitosan Blend Films

Blend films consisting of chlorophyllin dispersed in carboxymethyl chitin (CM-chitin) and carboxymethyl chitosan (CM-chitosan) were prepared by solution casting and characterized for their physical and electrical properties. Homogeneous films were obtained having chlorophyllin content up to 50 wt% in the CM-chitin matrix and 30 wt% in the CM-chitosan matrix. Agglomeration of chlorophyllin particles in the chlorophyllin/CM-chitosan blend was observed, when chlorophyllin content reached 50 wt%. The electrical conductivity of CM-chitosan was higher than that of CM-chitin by approximately 2 orders of magnitude due to the higher content of amino polar groups. The electrical conductivity increased with increasing chlorophyllin content. The addition of metal ion salts improved the electrical conductivity of the blend films.     

Chemosensitization of HT29 and HT29-5FU Cell Lines by a Combination of a Multi-Tyrosine Kinase Inhibitor and 5FU Downregulates ABCC1 and Inhibits PIK3CA in Light of Their Importance in Saudi Colorectal Cancer

Colorectal cancer (CRC) remains one of the main causes of death worldwide and in Saudi Arabia. The toxicity and the development of resistance against 5 fluorouracil 5FU pose increasing therapeutic difficulties, which necessitates the development of personalized drugs and drug combinations. Objectives: First, to determine the most important kinases and kinase pathways, and the amount of ABC transporters and KRAS in samples taken from Saudi CRC patients. Second, to investigate the chemosensitizing effect of LY294002 and HAA₂₀₂₀ and their combinations with 5FU on HT29, HT29-5FU, HCT116, and HCT116-5FU CRC cells, their effect on the three ABC transporters, cell cycle, and apoptosis, in light of the important kinase pathways resulting from the first part of this study. Methods: The PamChip^® peptide micro-array profiling was used to determine the level of kinase and targets in the Saudi CRC samples. Next, RT-PCR, MTT cytotoxicity, Western blotting, perturbation of cell cycle, annexin V, and immunofluorescence assays were used to investigate the effect on CRC, MRC5, and HUVEC cells. Results: The kinase activity profiling highlighted the importance of the PI3K/AKT, MAPK, and the growth factors pathways in the Saudi CRC samples. PIK3CA was the most overexpressed, and it was associated with increased level of mutated KRAS and the three ABC transporters, especially ABCC1 in the Saudi samples. Next, combining HAA₂₀₂₀ with 5FU exhibited the best synergistic and resistance-reversal effect in the four CRC cells, and the highest selectivity indices compared to MRC5 and HUVEC normal cells. Additionally, HAA₂₀₂₀ with 5FU exerted significant inhibition of ABCC1 in the four CRC cells, and inhibition of PIK3CA/AKT/MAPK7/ERK in HT29 and HT29-5FU cells. The combination also inhibited EGFR, increased the preG₁/S cell cycle phases, apoptosis, and caspase 8 in HT29 cells, while it increased the G₁ phase, p21/p27, and apoptosis in HT29-5FU cells. Conclusion: We have combined the PamChip kinase profiling of Saudi CRC samples with in vitro drug combination studies in four CRC cells, highlighting the importance of targeting PIK3CA and ABCC1 for Saudi CRC patients, especially given that the overexpression of PIK3CA mutations was previously linked with the lack of activity for the anti-EGFRs as first line treatment for CRC patients. The combination of HAA₂₀₂₀ and 5FU has selectively sensitized the four CRC cells to 5FU and could be further studied.     

Combination of 5-Fluorouracil with Photodynamic Therapy: Enhancement of Innate and Adaptive Immune Responses in a Murine Model of Actinic Keratosis†

We previously showed that a combination of differentiation-inducing agents (5-fluorouracil [5FU], vitamin D3 or methotrexate) and aminolevulinate-based photodynamic therapy (PDT) improves clinical responses by enhancing protoporphyrin IX (PpIX) photosensitizer levels and cell death. Here, we show that in addition to its previously known effects, 5FU enhances PDT-induced tumor-regressing immunity. Murine actinic keratoses were treated with topical 5FU or vehicle for 3 days prior to aminolevulinic acid application, followed by blue light illumination (~417 nm). Lesions were harvested for time-course analyses of innate immune cell recruitment into lesions, i.e. neutrophils (Ly6G+) and macrophages (F4/80+), which peaked at 72 h and 1 week post-PDT, respectively, and were greater in 5FU-treated lesions. Enhanced infiltration of activated T cells (CD3+) throughout the time course, and of cytotoxic T cells (CD8+) at 1–2 weeks post-PDT, also occurred in 5FU-treated lesions. 5FU pretreatment reduced the presence of cells expressing the immune checkpoint marker PD-1 at ~72 h post-PDT, favoring cytotoxic T cell activity. A combination of 5FU and PDT, each individually known to induce long-term tumor-targeting immune responses in addition to their more immediate effects on cancer cells, may synergize to provide better management of squamous precancers.     

Determination of 5-fluorouracil in serum microsamples by a high-performance liquid chromatographic method and its application to pharmacokinetic studies in rats

A high-performance liquid chromatographic method is described for the quantitative determination of 5-fluorouracil (5FU) in only 30 microL of serum. A reversed-phase C(18) column is used for the separation and analysis of 5FU. The mobile phase consists of methanol-20mmol/L phosphate buffer, pH 6.80 (5:95, v/v). The calibration curve for 5FU is linear over the range of 0.05-10 microg/mL. The extraction recoveries of 5FU and p-aminobenzoic acid (as an internal standard) are > 91.28% and 81.98%, respectively. The intra-day and inter-day coefficients of variation of 5FU are less than 6.29% and 9.57% at four different concentrations, respectively. The method is simple, sensitive, and reliable. It is superior to previous methods in that the sample volume needed is relatively small (just 30 microL of serum). Therefore, this method can be utilized to determine 5FU in rats for study in various fields, especially in pharmacokinetics, controlled-release, and combination therapy with 5FU, etc. In this study, it is successfully applied to pharmacokinetic studies of 5FU after administrations of an intravenous bolus dose (25 mg/kg) and two oral doses (25 and 75 mg/kg) in rats.     

LC‐MS/MS method for simultaneous analysis of uracil, 5,6‐dihydrouracil, 5‐fluorouracil and 5‐fluoro‐5,6‐dihydrouracil in human plasma for therapeutic drug monitoring and toxicity prediction in cancer patients

The chemotherapeutic drug 5‐fluorouracil (5‐FU) is widely used for treating solid tumors. Response to 5‐FU treatment is variable with 10–30% of patients experiencing serious toxicity partly explained by reduced activity of dihydropyrimidine dehydrogenase (DPD). DPD converts endogenous uracil (U) into 5,6‐dihydrouracil (UH₂), and analogously, 5‐FU into 5‐fluoro‐5,6‐dihydrouracil (5‐FUH₂). Combined quantification of U and UH₂ with 5‐FU and 5‐FUH₂ may provide a pre‐therapeutic assessment of DPD activity and further guide drug dosing during therapy. Here, we report the development of a liquid chromatography–tandem mass spectrometry assay for simultaneous quantification of U, UH₂, 5‐FU and 5‐FUH₂ in human plasma. Samples were prepared by liquid–liquid extraction with 10:1 ethyl acetate‐2‐propanol (v/v). The evaporated samples were reconstituted in 0.1% formic acid and 10 μL aliquots were injected into the HPLC system. Analyte separation was achieved on an Atlantis dC₁₈ column with a mobile phase consisting of 1.0 mm ammonium acetate, 0.5 mm formic acid and 3.3% methanol. Positively ionized analytes were detected by multiple reaction monitoring. The analytical response was linear in the range 0.01–10 μm for U, 0.1–10 μm for UH₂, 0.1–75 μm for 5‐FU and 0.75–75 μm for 5‐FUH₂, covering the expected concentration ranges in plasma. The method was validated following the FDA guidelines and applied to clinical samples obtained from ten 5‐FU‐treated colorectal cancer patients. The present method merges the analysis of 5‐FU pharmacokinetics and DPD activity into a single assay representing a valuable tool to improve the efficacy and safety of 5‐FU‐based chemotherapy. Copyright © 2012 John Wiley & Sons, Ltd.     

Effects of American ginseng on pharmacokinetics of 5‐fluorouracil in rats

The pharmacokinetics of 5‐fluorouracil (5‐FU) in combination with or without American ginseng (seven‐consecutive days oral dose) in rats were evaluated using liquid chromatography–electrospray ionization–mass spectrometry (LC‐MS). Chromatographic separation was performed on a reverse LC column within a total run time of 6.5 min, which allowed for a relatively quick analysis. The limit of quantification for 5‐FU was 15 ng/mL and this method was linear over 15–50,000 ng/mL. This method supported stabilizing determination of the plasma concentration of 5‐FU over a period of 24 h. Precision both interday and intraday (coefficient of variation) was within 14% and accuracy (relative error) ranged from ?5 to 14%. In view of the observed pharmacokinetic parameters, including maximum concentration, time to maximum concentration, area under the concentration–time curve (AUC), mean residence time, elimination half‐life and clearance, our results showed no significant differences in all of the pharmacokinetic parameters between the ginseng co‐treated group and 5‐FU alone group. Some increase in AUC was observed in 5‐FU plus ginseng group; however, the difference did not reach statistical significance compared with 5‐FU alone. It appeared that American ginseng administration did not significantly alter the kinetics of 5‐FU. More studies are still needed to confirm our results. Copyright © 2014 John Wiley & Sons, Ltd.     

A new high-performance liquid chromatographic/electrospray ionization tandem mass spectrometric method for the simultaneous determination of cyclophosphamide, methotrexate and 5-fluorouracil as markers of surface contamination for occupational exposure monitoring

A new high-performance liquid chromatographic/electrospray ionization tandem mass spectrometric (HPLC/ESI-MS/MS) method was developed for the simultaneous quantification of 5-fluorouracil (5FU), methotrexate (MTX) and cyclophosphamide (CP) in environmental samples. These compounds, commonly used in the treatment of cancer, are recognized as genotoxic. In order to estimate the occupational exposure of hospital personnel handling these drugs, wipe samples were taken from the working surfaces and directly analyzed (with trophosphamide as internal standard) using a reversed-phase capillary column and MS/MS detection. This is the first HPLC/MS/MS method for the simultaneous determination of 5FU, MTX and CP. The present method offers high sensitivity, with detection limits of 1.1 microg l(-1) for MTX and CP and 33.3 microg l(-1) for 5FU, avoiding any sample preconcentration procedure. Rapidity, specificity, high accuracy (mean values between 92.4 and 99.9%) and precision (mean RSD values between 3.4 and 12.1%) make the method suitable for the routine determination of these three antineoplastic drugs.     

Effect of Laser Phototherapy on Enzymatic Activity of Salivary Glands of Hamsters Treated with 5‐Fluorouracil

The chemotherapeutic agent 5‐Fluorouracil (5‐FU) can induce salivary gland hypofunction (SGH); however, previous studies did not reach final conclusions on the influence of this drug on glandular tissue. Thus, the aim of this study was to investigate the effect of 5‐FU on submandibular (SMs) and sublingual glands (SLs), as well as, the effect of laser phototherapy (LPT) on SGH induced by 5‐FU. Eighty‐five hamsters were divided into three groups: control (C), chemotherapy (CT) and laser (L), and the SGH was induced by two injections of 5‐FU in groups CT and L. The irradiation was performed using a diode (λ780 nm/20 mW/5 J cm^?2/0.2 J and 10 s per point/spot size of 0.04 cm²) and applied daily. On the euthanasia day, SMs and SLs were removed and biochemical analyses were carried out. The lactate dehydrogenase activity was increased in group CT when compared with group C for SLs and SMs (P < 0.05). In addition, the peroxidase and catalase activities were increased and superoxide dismutase was decreased by 5‐FU (P < 0.05). However, LPT appears to be a protective mechanism against oxidative stress, tending to alter the activity of these antioxidant enzymes, suggesting LPT as a promising therapy to modulate the 5‐FU harmful effect.     

FTIR and circular dichroism spectroscopic study of interaction of 5-fluorouracil with DNA

5-Fluorouracil (5FU) is an anticancer chemotherapeutic drug which exerts cytotoxic effect by inhibiting cellular DNA replication. In the present study, we explore the binding of 5FU with DNA and resulting structural and conformational changes on DNA duplex. UV-visible, Fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopic techniques were employed to explore these interactions. A constant concentration of calf thymus DNA was incubated with varying concentrations of 5FU. UV-visible and FTIR spectroscopic results revealed that intercalation is the primary mode of interaction between 5FU and nitrogenous bases of the nucleic acid. The binding constant was found to be 9.7×10(4); which is indicative of moderate type of interaction between 5FU and DNA duplex. It was also observed that 5FU intercalates slightly more between AT base pairs compared to GC pairs. FTIR and circular dichroism spectroscopic results revealed that 5FU disturbs native B-conformation of DNA though, DNA remains in its B conformation even at higher concentrations of 5FU.     

A simultaneous determination method for 5‐fluorouracil and its metabolites in human plasma with linear range adjusted by in‐source collision‐induced dissociation using hydrophilic interaction liquid chromatography–electrospray ionization–tandem mass spectrometry

We applied a new technique for quantitative linear range shift using in‐source collision‐induced dissociation (CID) to complex biological fluids to demonstrate its utility. The technique was used in a simultaneous quantitative determination method of 5‐fluorouracil (5‐FU), an anticancer drug for various solid tumors, and its metabolites in human plasma by liquid chromatography–electrospray ionization–tandem mass spectrometry (LC/ESI‐MS/MS). To control adverse effects after administration of 5‐FU, it is important to monitor the plasma concentration of 5‐FU and its metabolites; however, no simultaneous determination method has yet been reported because of vastly different physical and chemical properties of compounds. We developed a new analytical method for simultaneously determining 5‐FU and its metabolites in human plasma by LC/ESI‐MS/MS coupled with the technique for quantitative linear range shift using in‐source CID. Hydrophilic interaction liquid chromatography using a stationary phase with zwitterionic functional groups, phosphorylcholine, was suitable for separation of 5‐FU from its nucleoside and interfering endogenous materials. The addition of glycerin into acetonitrile‐rich eluent after LC separation improved the ESI‐MS response of high polar analytes. Based on the validation results, linear range shifts by in‐source CID is the reliable technique even with complex biological samples such as plasma. Copyright © 2016 John Wiley & Sons Ltd.     

Fabrication,rheological analysis,and in vitro characterization of in situ chemically cross‐linkable thermogels as controlled and prolonged drug depot for localized and systemic delivery

5‐Fluorouracil (5‐FU) is widely used against many types of solid cancer in clinics. However, because of its limitations such as short half‐life, poor oral absorption and rapid clearance by dihydropyrimidine dehydrogenase have limited its applications. In current study, new in situ chemically grafted thermogels for prolonged drug release are formed on the basis of poloxamer 407 (PF127) and carboxymethyl chitosan (CMCS) using glutaraldehyde as cross‐linking agent. The phase transition from sol to gel state at body temperature was confirmed by tube titling, rheological analysis, and optical transmittance determinations. Swelling and drug release experiments conducted at various pH and temperature demonstrated that developed formulations are thermoresponsive with maximum swelling and release below critical gelation temperature (CGT) (pH 7.4, 25°C). Cells growth inhibition study confirmed the biocompatibility of thermogels against L929 cell lines. Methyl thiazolyl tetrazolium (MTT) assay confirmed that 5‐FU–loaded thermogels have the potential to cause cells death against HeLa and MCF‐7 cancer lines. The IC₅₀ values calculated for pure 5‐FU solution (27 ± 0.81 μg/mL for HeLa and 24 ± 0.58 μg/mL for MCF‐7) were found higher in comparison with 5‐FU–loaded thermogels, against HeLa (17 ± 0.39 μg/mL) and MCF‐7 (14 ± 0.67 μg/mL). Fourier transform infrared (FTIR) confirmed the new structure formation and chemical grafting between PF127 and CMCS. Thermogravimetric (TG) and differential scanning calorimetry (DSC) analyses proved the phase transition around physiologic temperature range, while scanning electron microscopy (SEM) analysis displayed the presence of connected pores in the cross section of thermogels facilitating the uptake of solvents and drug particles. Altogether, results concluded that developed chemically grafted thermogels can be used in vivo for prolonged drug release after subcutaneous administration.