Metformin hydrochloride

Manufactured by Tokyo Chemical Industry

Sourced in Japan

Metformin hydrochloride is a white, crystalline powder used as a laboratory reagent. It is the hydrochloride salt of the antidiabetic drug metformin.

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Lab products found in correlation

8 protocols using metformin hydrochloride

Metabolic Modulators in Cell Assays

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Metformin hydrochloride (Tokyo Chemical Industry), 2-deoxy-D-glucose (Sigma-Aldrich), Etomoxir (Selleck Chemicals), ethyl sulfide (BPTES, Sigma-Aldrich), Chloroquine hydrochloride (Sigma-Aldrich), UK5099 (Sigma-Aldrich), epigallocatechin gallate (Tokyo Chemical Industry), 3MPA (Sigma-Aldrich), MitoTempo (Sigma-Aldrich), Compound C (Sigma-Aldrich) were purchased.

Chao R., Nishida M., Yamashita N., Tokumasu M., Zhao W., Kudo I, & Udono H. (2022). Nutrient Condition in the Microenvironment Determines Essential Metabolisms of CD8+ T Cells for Enhanced IFNγ Production by Metformin. Frontiers in Immunology, 13, 864225.

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Inflammatory Cytokine Signaling Modulation

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Human recombinant TNF-α, IL-17A and IL-1β were purchased from PeproTech (Rocky Hill, NJ, USA). Dimethyl sulfoxide (DMSO) was purchased from Nacalai Tesque (Kyoto, Japan). Sirtinol (Enzo Life Science, Farmingdale, NY, USA) and dorsomorphin (Abcam, Cambridge, UK) were dissolved in DMSO and stored at −30 °C until used in the experiments. Metformin (Metformin hydrochloride, Tokyo Chemical Industry, Tokyo, Japan) was dissolved in medium and filtrated by 0.22 µm polyvinylidene difluoride (PVDF) membrane (Merck Millipore, Burlington, MA, USA). Anti-human pro-IL-1β monoclonal rabbit antibody, anti-human NLRP3 monoclonal rabbit antibody, anti-human caspase-1 monoclonal rabbit antibody, anti-human β-actin monoclonal mouse antibody, anti-phosphorylated AMPKα rabbit monoclonal antibody (Thr172), anti-AMPKα rabbit monoclonal antibody, anti-human S100A9 monoclonal rabbit antibody (Cell Signaling Technology, Danvers, MA, USA), anti-human IL-36γ monoclonal mouse antibody, anti-human S100A7 monoclonal mouse antibody and anti-human S100A8 monoclonal rabbit antibody (Abcam) were used for western blotting.

Tsuji G., Hashimoto-Hachiya A., Yen V.H., Takemura M., Yumine A., Furue K., Furue M, & Nakahara T. (2020). Metformin inhibits IL-1β secretion via impairment of NLRP3 inflammasome in keratinocytes: implications for preventing the development of psoriasis. Cell Death Discovery, 6, 11.

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Extraction and Purification of Arctigenin

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Powdered GSE was extracted from dried burdock sprouts using hydrated ethanol. The extract
was found to contain 13.7% AG as quantified using high-performance liquid chromatography
(HPLC). Pure AG (>95.0%) was then obtained from the GSE using the method described by
Liu et al. [19 (link)]. GSE and AG were
produced by our company, Kracie Holdings, Ltd. (GSE, lot no. 170927: AG, lot no. 190422).
Metformin hydrochloride was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo,
Japan).

WATANABE S., OHNO A., YOMODA S, & INAMASU S. (2022). Arctigenin-containing burdock sprout extract prevents obesity in association with modulation of the gut microbiota in mice. Bioscience of Microbiota, Food and Health, 42(1), 49-55.

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Metformin and Cell Depletion in Osteosarcoma

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Mice were intra-dermally inoculated in the back flank (right side) with 4 × 10⁵ K7M2neo cells using a 27-gauge needle. Subsequently, mice were orally administered 5 mg ml⁻¹ of metformin hydrochloride (Tokyo Chemical Industry Co., Ltd, Japan) dissolved in free drinking water. To deplete T cells in vivo, 50 µl ascites formed with antibodies against CD4 and CD8 molecules was diluted in 150 µl phosphate-buffered saline (PBS) and injected into mice from day 7 at 4-day intervals, as described previously (30 (link)). To deplete CD11b⁺ cells in vivo, 100 µg anti-CD11b antibody (5C6 clone 1, BioGate Co., Ltd, Gifu, Japan) was injected in 5-day intervals throughout the experiments. The long (a) and short (b) tumor diameters were measured using Vernier calipers, while tumor volume (V) was calculated according to the following equation: V = a × b²/2.

Uehara T., Eikawa S., Nishida M., Kunisada Y., Yoshida A., Fujiwara T., Kunisada T., Ozaki T, & Udono H. (2018). Metformin induces CD11b+-cell-mediated growth inhibition of an osteosarcoma: implications for metabolic reprogramming of myeloid cells and anti-tumor effects. International Immunology, 31(4), 187-198.

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Fabrication of Metformin Microstirrer Pills

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Microstirring pills were fabricated by triturating and mixing lactose (Spectrum Chemical MGF Corp.) and maltose (Spectrum Chemical MGF Corp.) at a 60%/40% ratio. Once this mixture was homogeneous, microstirrers (10% of the total mixture weight) were incorporated, followed by homogenization in a mortar. No changes in the microstirrer structure were observed during this trituration and fabrication process. Metformin hydrochloride (Tokyo Chemical Industry) was also added at this step. In order to obtain a paste-like consistency, an ethanol/water wetting solution (75%/25%) was added to the matrix mixture. Subsequently, the paste was transferred to a cavity plate and all the cavities were filled with the paste by applying enough pressure to guarantee a tight packing. Right after filling each of the cavities, the plate was lowered onto the peg plate until the wet pills were ejected. Lastly, all the pills were allowed to dry and harden over the peg plate for 2 h at 65 °C. Pills without microstirrers were fabricated following the same protocol. After this entire process, the pills were enterically coated with Eudragit L-100 (Evonik Corporations GmbH), 8% v/v, by a dip coating technique.

Ziesche R., Roth M., Papakonstantinou E., Nauck M., Hörl W.H., Kashgarian M, & Block L.H. (1994). A granulocyte inhibitory protein overexpressed in chronic renal disease regulates expression of interleukin 6 and interleukin 8.. Proceedings of the National Academy of Sciences of the United States of America, 91(1), 301-305.

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Investigating NF-κB Signaling Pathway

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Anti-murine NF-κB p65 monoclonal rabbit antibody (Abcam, Cambridge, UK), anti-murine NF-κB p65 (phospho Ser536) polyclonal rabbit antibody (Abcam), anti-murine IκBζ (protein corded by NFKBIZ gene) polyclonal rabbit antibody, and anti-murine β-actin monoclonal mouse antibody (Cell Signaling Technology, Danvers, MA, USA) were used for Western blotting. Dimethyl Sulfoxide (DMSO) was purchased from Nacalai Tesque, Inc. (Kyoto, Japan). Metformin hydrochloride and BAY 11-7082 were obtained from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Murine recombinant IL-36γ was obtained from R&D Systems (Minneapolis, MN, USA).

Matsuda-Taniguchi T., Takemura M., Nakahara T., Hashimoto-Hachiya A., Takai-Yumine A., Furue M, & Tsuji G. (2021). The Antidiabetic Agent Metformin Inhibits IL-23 Production in Murine Bone-Marrow-Derived Dendritic Cells. Journal of Clinical Medicine, 10(23), 5610.

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Fabrication and Enteric Coating of Microstirring Pills

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Microstirring pills were fabricated by triturating and mixing lactose (Spectrum Chemical MGF Corp.) and maltose (Spectrum Chemical MGF Corp.) at a 60%/40% ratio. Once this mixture was homogeneous, microstirrers (10% of the total mixture weight) were incorporated, followed by homogenization in a mortar. No changes in the microstirrer structure were observed during this trituration and fabrication process. Metformin hydrochloride (Tokyo Chemical Industry) was also added at this step. In order to obtain a paste-like consistency, an ethanol/water wetting solution (75%/25%) was added to the matrix mixture. Subsequently, the paste was transferred to a cavity plate and all the cavities were filled with the paste by applying enough pressure to guarantee a tight packing. Right after filling each of the cavities, the plate was lowered onto the peg plate until the wet pills were ejected. Lastly, all the pills were allowed to dry and harden over the peg plate for 2 h at 65 °C.
Pills without microstirrers were fabricated following the same protocol. After this entire process, the pills were enterically coated with Eudragit L-100 (Evonik Corporations GmbH), 8% v/v, by a dip coating technique.

Mundaca-Uribe R., Holay M., Abbas A., Askarinam N., Sage-Sepulveda J.S., Kubiatowicz L., Fang R.H., Zhang L, & Wang J. (2023). A Microstirring Oral Pill for Improving the Glucose-Lowering Effect of Metformin. ACS nano, 17(10).

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Pharmaceutical Solid Dispersion Formulation

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Metformin hydrochloride (MET) was purchased from Tokyo Chemical Industry UK Ltd.
(Oxford, UK) and sitagliptin phosphate (SIT) was purchased from Kemprotec (Carnforth, UK).
Three hydroxypropyl cellulose (HPC) grades were studied as carrier polymers/meltable binders and their characteristics are detailed in Table I. HPC-A was obtained from Ashland (Covington, Kentucky, USA), HPC-S was received from Sigma-Aldrich (Dorset, UK) and L-HPC was obtained from Shin-Etsu (Wiesbaden, Germany). Polyethylene glycol (3350 g/ mol) was purchased from Sigma-Aldrich (Dorset, UK). Microcrystalline cellulose, Avicel PH102, was received from IMCD Ltd. (Sutton, UK). Deionised water was obtained using an Elix 3 connected to a Synergy® UV water purification system (Millipore, UK). All other chemicals were obtained from Sigma-Aldrich (Dorset, UK) and were of analytical grade or equivalent and were used without any further treatment.

Kelleher J.F., Madi A.M., Gilvary G.C., Tian Y.W., Li S., Almajaan A., Loys Z.S., Jones D.S., Andrews G.P, & Healy A.M. (2019). Metformin Hydrochloride and Sitagliptin Phosphate Fixed-Dose Combination Product Prepared Using Melt Granulation Continuous Processing Technology. AAPS PharmSciTech, 21(1).

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