Chemical characterizations for article Gemcitabine release kinetics and biological activity of HPMA copolymer nanomedicines

--------------------------------------------------------------------------------------------------------- Chemical characterizations for article Gemcitabine release kinetics and biological activity of HPMA copolymer nanomedicines --------------------------------------------------------------------------------------------------------- ReadMe version: 1.0 (2026-11-03) Dataset version: 1.0 (2026-11-03) This Dataset DOI: 10.5281/zenodo.23950053 Related journal publication DOI: 10.1016/j.jconrel.2026.115396 Related dataset DOI: 10.48700/3nvnc-kk729 (Contains the biological research data for the same paper) -------------------------------------------------------------------- CONTACT -------------------------------------------------------------------- Tomáš Etrych | [e-mail] ORCID: 0000-0001-5908-5182 Institute of Macromolecular Chemistry, Czech Academy of Sciences Heyrovského náměstí 1888/2, Prague 6 – Břevnov, 162 00, Czech Republic ______Creators______ * Robert Pola (ORCID: 0000-0003-0025-2016; Institute of Macromolecular Chemistry CAS) — Writing – original draft; Visualization; Methodology; Investigation; Data curation. * Petr Kašpárek (Institute of Macromolecular Chemistry CAS) — Investigation; Data curation. * Alžběta Bulková (ORCID: 0009-0004-7687-6014; Institute of Macromolecular Chemistry CAS) — Investigation; Data curation. * Tomáš Etrych (ORCID: 0000-0001-5908-5182; Institute of Macromolecular Chemistry CAS) — Writing – review & editing; Supervision; Project administration; Funding acquisition; Conceptualization. -------------------------------------------------------------------- DATA AVAILABILITY AND ACCESS INSTRUCTIONS -------------------------------------------------------------------- The dataset is openly accessible under the DOI listed above. The license and terms of reuse are shown in the following chapter below. -------------------------------------------------------------------- LICENSE -------------------------------------------------------------------- ______ReadMe file license______ ReadMe by Robert Pola is licensed under CC BY 4.0 License information: https://creativecommons.org/licenses/by/4.0/ ______Dataset license______ Chemical characterizations for article Gemcitabine release kinetics and biological activity of HPMA copolymer nanomedicines by Pola, Kašpárek, Bulková and Etrych is licensed under CC BY 4.0 License information: https://creativecommons.org/licenses/by/4.0/ -------------------------------------------------------------------- DESCRIPTION AND METHODOLOGY -------------------------------------------------------------------- ______About the dataset______ This study examined how the chemical structure of linkers joining gemcitabine (Gem) to N-(2-hydroxypropyl) methacrylamide (HPMA) copolymers affects drug-release kinetics, pharmacokinetics, antitumor efficacy and immunomodulatory activity. Four conjugates (P-Gem1: GFLG linker; P-Gem2: β-alanine; P-Gem3: 5-aminopentanoic acid/Apt; P-Gem4: 6-aminohexanoic acid/Ahx) were synthesized and characterized. Data cover physicochemical characterization, drug release in PBS at pH 7.4, 6.7 and 6.0 and mouse sera, in vitro cancer-cell assays, toxicity, antitumor activity in murine and human xenograft tumor models, pharmacokinetic bioassays, tumor gene expression, apoptosis, and immune-cell profiling. ______Sample preparation______ HPMA copolymer precursors Prec1–4 were synthesized via RAFT copolymerization using HPMA and TT-functionalized linker monomers (88:12 molar feed) followed by removal of dithiobenzoate end groups. Gem was coupled to the reactive precursors in pyridine at 50 °C for 44 h, and products were purified by precipitation, Sephadex G-25 chromatography and freeze-drying. Solutions of P-Gem conjugates were incubated in phosphate buffer or murine sera at 37 °C for release analysis. Further detail can be found in the publication 10.1016/j.jconrel.2026.115396 ______Methods of data collection______ * Chemical/physical characterization: **HPLC Conjugates & Monomenrs (Chromolith Performance RP- 18e column, 100 ×4.6 mm, Merck, Germany, UV diode-array, Shimadzu, Japan). Details of the methods can be found in the individual files’ metadata. **HPLC Release of conjugates incubated in phosphate buffer (pH 7.4, 6.7, 6.0) or mouse serum at 37 °C; release experiments reported as three independent replicates. (Chromolith Performance RP- 18e column, 100 ×4.6 mm, Merck, Germany, UV diode-array, Shimadzu, Japan). Details of the methods can be found in the individual files’ metadata. **SEC) on an HPLC system (Shimadzu, Japan) equipped with: UV, differential refractive index and multi-angle light scattering DAWN Heleos II detectors (Wyatt Technology Corp., USA). The analysis was performed using a TSK 3000 SWXL column (Tosoh Bioscience, Japan). Details of the methods can be found in the individual files’ metadata. ______Methods of data processing______ HPLC-based purity of monomers and conjugates, quantification of release, SEC-derived molar masses/dispersities. Details can be found in the associated journal publication. -------------------------------------------------------------------- DATASET STRUCTURE -------------------------------------------------------------------- ├── 000_ReadMe.txt ├── 001_HPLC │ ├── Conjugates │ │ ├── P-Gem1.lcd │ │ ├── (… 2x) │ │ └── P-Gem4.lcd │ ├── Monomers │ │ ├── Ma-Acap-TT.lcd │ │ ├── (… 2x) │ │ └── Ma-GFLG-TT.lcd │ └── Release │ ├── Buffers │ │ ├── P-Gem1 │ │ │ ├── 6.0 - 0day.lcd │ │ │ ├── (… 19x) │ │ │ └── 7.4 - 7days.lcd │ │ ├── P-Gem2 │ │ │ ├── 6.0 - 0day.lcd │ │ │ ├── (… 19x) │ │ │ └── 7.4 - 7days.lcd │ │ ├── P-Gem3 │ │ │ ├── 6.0 - 0day.lcd │ │ │ ├── (… 19x) │ │ │ └── 7.4 - 7days.lcd │ │ └── P-Gem4 │ │ ├── 6.0 - 0day.lcd │ │ ├── (… 19x) │ │ └── 7.4 - 7days.lcd │ └── Serum │ ├── P-Gem1 │ │ ├── Fresh BALB-c │ │ │ ├── 0hrs.lcd │ │ │ ├── (…5x) │ │ │ └── 6hrs.lcd │ │ ├── Fresh C57BL-6 │ │ │ ├── 0hrs.lcd │ │ │ ├── (…5x) │ │ │ └── 6hrs.lcd │ │ └── Frozen serum │ │ ├── 0hrs.lcd │ │ ├── (…5x) │ │ └── 6hrs.lcd │ ├── P-Gem2 │ │ ├── Fresh BALB-c │ │ │ ├── 0hrs.lcd │ │ │ ├── (…5x) │ │ │ └── 6hrs.lcd │ │ ├── Fresh C57BL-6 │ │ │ ├── 0hrs.lcd │ │ │ ├── (…5x) │ │ │ └── 6hrs.lcd │ │ └── Frozen serum │ │ ├── 0hrs.lcd │ │ ├── (…5x) │ │ └── 6hrs.lcd │ ├── P-Gem3 │ │ ├── Fresh BALB-c │ │ │ ├── 0hrs.lcd │ │ │ ├── (…5x) │ │ │ └── 6hrs.lcd │ │ ├── Fresh C57BL-6 │ │ │ ├── 0hrs.lcd │ │ │ ├── (…5x) │ │ │ └── 6hrs.lcd │ │ └── Frozen serum │ │ ├── 0hrs.lcd │ │ ├── (…5x) │ │ └── 6hrs.lcd │ └── P-Gem4 │ ├── Fresh BALB-c │ │ ├── 0hrs.lcd │ │ ├── (…5x) │ │ └── 6hrs.lcd │ ├── Fresh C57BL-6 │ │ ├── 0hrs.lcd │ │ ├── (…5x) │ │ └── 6hrs.lcd │ └── Frozen serum │ ├── 0hrs.lcd │ ├── (…5x) │ └── 6hrs.lcd └── 002_SEC ├── P-Gem1.vaf ├── P-Gem2.vaf ├── P-Gem3.vaf ├── P-Gem4.vaf ├── Prec1.vaf ├── Prec2.vaf ├── Prec3.vaf └── Prec4.vaf -------------------------------------------------------------------- FILENAME STRUCTURE -------------------------------------------------------------------- ___001_HPLC___ *001_HPLC /Conjugates: P-Gem1, P-Gem2, P-Gem3, and P-Gem4 (HPMA copolymer–gemcitabine conjugates). e.g. P-Gem1.lcd *001_HPLC/Monomers: Ma-GFLG-TT, Ma-bAla-TT, Ma-Apt-TT, and Ma-Ahx-TT (thiazolidine-2-thione-containing monomers used for polymer precursor synthesis) e.g. Ma-GFLG-TT.lcd, * 001_HPLC/release data HPLC data documenting gemcitabine release from polymer conjugates P-Gem1–4 under different incubation conditions are organized into separate subfolders for each conjugate (P-Gem1 to P-Gem4). Buffer release data contain the pH value (7.4, 6.7, or 6.0) in the file name, whereas serum release data are further organized into subfolders according to the serum type (Fresh BALB-c, Fresh C57BL-6, or Frozen serum). File names indicate the incubation time expressed in hours (xhrs) or days (xday or xdays), where x represents the corresponding numerical value. All HPLC data files are stored in the .lcd format. Filename convention - Buffers: File names contain the incubation pH and time. - Serum: File names indicate the incubation time; serum type is specified by the subfolder name. ___002_SEC___ *Prec[N].vaf for polymer precursors and *P-Gem[N].vaf for gemcitabine conjugates N = 1, 2, 3 or 4. e.g. Prec1.vaf -------------------------------------------------------------------- FILE TYPES & FORMATS, SOFTWARE TO OPEN, DIMENSIONS & UNITS -------------------------------------------------------------------- ___SEC___ Molecular weight Mw (g/mol) and dispersity Đ (dimensionless). Original format: .vaf SW: Astra (V 5.3.4.20) ___HPLC__ Gem content: wt%; linker TT content: mol%. Drug release: time (days), conjugated/released Gem (%). Original format: .lcd SW: LabSolutions (version: 1.25 SP2) -------------------------------------------------------------------- FUNDING / ACKNOWLEDGMENT -------------------------------------------------------------------- *Czech Health Research Council — NW25-03-00150 (M. Kovář and T. Etrych). *National Institute for Cancer Research (Programme EXCELES), project LX22NPO5102; funded by the European Union — Next Generation EU. *Institutional Research Concept RVO 61388971.