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Doxycycline hyclate [24390-14-5]

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Cat# HY-N0565B-5g

Size : 5g

Brand : MedChemExpress

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Description

Doxycycline hyclate, an antibiotic, is an orally active and broad-spectrum metalloproteinase (MMP) inhibitor. Doxycycline hyclate shows antibacterial activity and anti-cancer cell proliferation activity. Doxycycline hyclate can be used to construct gene expression regulation models^[1]^[2]^[3]^[4]^[5].

IC₅₀ & Target

Tetracycline

Cellular Effect

Cell Line	Type	Value	Description	References
HepG2	CC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells assessed as reduction in cell viability measured after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells assessed as reduction in cell viability measured after 72 hrs by MTT assay	[PMID: 34364164]
HepG2	CC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells assessed as reduction in cell viability after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells assessed as reduction in cell viability after 72 hrs by MTT assay	[PMID: 27654395]
HepG2	CC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells measured after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells measured after 72 hrs by MTT assay	[PMID: 27155463]
HepG2	CC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells assessed as cell viability after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells assessed as cell viability after 72 hrs by MTT assay	[PMID: 25791675]
HepG2	CC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells incubated for 72 hrs by MTT assay Cytotoxicity against human HepG2 cells incubated for 72 hrs by MTT assay	[PMID: 25282267]
HepG2	CC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay	[PMID: 24946216]
HepG2	CC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay	[PMID: 22889559]
HepG2	CC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay	[PMID: 21852132]
HepG2	CC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay	[PMID: 21741131]
HepG2	IC₅₀	20 μM Compound: doxycycline	Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay	[PMID: 19926173]
HepG2	IC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells after 72 hrs by MTT assay	[PMID: 19482476]
HT-1080	IC₅₀	21.4 μM Compound: 7	Inhibition of cell invasion in human HT-1080 cells for 24 hrs by Calcein AM staining based fluorometric analysis Inhibition of cell invasion in human HT-1080 cells for 24 hrs by Calcein AM staining based fluorometric analysis	[PMID: 35969157]
K562	IC₅₀	15 μM Compound: doxycycline	Cytotoxicity against human K562 cells after 72 hrs by flow cytometry Cytotoxicity against human K562 cells after 72 hrs by flow cytometry	[PMID: 19926173]
K562	IC₅₀	15 μM Compound: Doxycycline	Cytotoxicity against human K562 cells after 72 hrs by flow cytometry Cytotoxicity against human K562 cells after 72 hrs by flow cytometry	[PMID: 19482476]
THP-1	IC₅₀	20 μM Compound: Doxycycline	Cytotoxicity against human THP1 cells after 72 hrs by propidium iodide staining-based flow cytometry Cytotoxicity against human THP1 cells after 72 hrs by propidium iodide staining-based flow cytometry	[PMID: 19748781]

In Vitro

Doxycycline hyclate (0.01-10 μg/mL, 4 d) affects growth of glioma cells only under high concentrations^[2].
Doxycycline hyclate (0.01-10 μg/mL, 24 h) decreases MT-CO1 protein content with concentrations of 1 μg/mL and higher in SVG cells^[2].
Doxycycline hyclate (100 ng/mL, 1 μg/mL; 24 h) reduces proliferation of human cell lines^[4].
Doxycycline hyclate (0-250 μM, 72 h) inhibits cell viability of breast cancer cells ^[5].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Viability Assay^[2]

Cell Line:	LNT-229, G55, and U343 glioma cells
Concentration:	0.01, 0.1, 1 or 10 µg/mL
Incubation Time:	4 days
Result:	Affected growth of glioma cells only under high concentrations (10 µg/mL).

Cell Viability Assay^[2]

Cell Line:	SVG cells
Concentration:	0.01, 0.1, 1 or 10 µg/mL
Incubation Time:	24 hours
Result:	Decreaseed MT-CO1 protein content with concentrations of 1 µg/mL and higher.

Cell Proliferation Assay^[4]

Cell Line:	MCF 12A, 293T cells
Concentration:	100 ng/mL, 1 µg/mL
Incubation Time:	96 hours
Result:	Caused reduced proliferation of MCF 12A and 293T cells at 1 µg/mL.

Cell Viability Assay^[5]

Cell Line:	MCF-7, MDA-MB-468 cells
Concentration:	0-250 μM
Incubation Time:	72 hours
Result:	Inhibited breast cancer cells in a dose-dependent manner with IC₅₀ values for MCF-7 and MDA-MB-468 of 11.39 μM and 7.13 μM respectively.

In Vivo

Doxycycline (oral gavage; 200 or 800 mg/kg; once daily; 3 months) hyclate reduces MMP-9 activity in untreated HT mice in a dose-dependent manner^[3].
Doxycycline hyclate and Tetracycline (HY-A0107), act systemically after absorption from the upper gastrointestinal tract. The main advantage of Doxycycline hyclate over Tetracycline is its longer activity, and it can be taken twice or once a day. The peak concentration of both agents is similar, but in the case of Doxycycline the time to peak concentration is shorter, and half life is significantly longer^[6].

Doxycycline (Dox) hyclate is often used as an inducer in molecular biology studies to induce gene expression. In cells or model animals that have constructed tetracycline induced expression systems (Tet-On/Tet-Off systems), the expression of target genes can be precisely controlled by adding or removing Dox^[7]^[8]^[9]^[10].
Dose reference for Dox induction^[7]^[8]:
(1) Model animal: male Sprague-Dawley rats
Tet regulatory system: 20-3000 ppm of Dox is supplied in diet.
(2) Model animal: Cags mice
Tet regulatory system: 625 ppm of Dox is supplied in diet.

Induction of Modeling ON-OFF System (Gene expression regulation)^[6]^[7]^[8]

Background

Doxycycline hyclate is often used as an inducer in molecular biology research to induce gene expression. In cells or model animals that have constructed a Tetracycline (Tet; HY-A0107) inducible expression (Tet-ON/Tet-OFF) system, the expression of the target gene can be precisely controlled by adding or removing Doxycycline hyclate. Doxycycline hyclate can act as an inhibitor of transcriptional activation in the Tetracycline (Tc)-controlled transactivation (tTA) system, and as an inducer of transcriptional activation in the "reverse tTA' system.
Doxycycline and Tetracycline both act systemically after being absorbed by the upper gastrointestinal tract. In comparison, the main advantage of Doxycycline is that it has a longer activity and can be taken twice or once a day. Although the peak concentrations of the two are similar, Doxycycline takes a shorter time to reach peak concentration and has a significantly longer half-life.

Specific Modeling Methods

Rat^[8]: Sprague-Dawley rats • male • adult middle-aged (12-month-old)
Administration: (for GDNF as targeted gene) 3g/kg (Doxycycline; dietary with regular food) • po • once daily for 6 days

Note

(1) Recommend use the recombinant adeno-associated virus (rAAV)-based bicistronic tetracycline (tet)-OFF construct was used for dynamic control of GDNF (target gene) expression during long-term expression^[7].
(2) 3 g/kg dietary DOX produced DOX serum levels equivalent to 1mg/ml DOX in drinking water.

Modeling Indicators

Molecular changes: The expression level of the target gene decreases.

Phenotype changes: The positively correlated phenotype corresponding to the target gene is alleviated.

Opposite Product(s): /

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model:	6-month-old female Heterozygous Col3a1-deficient (HT) mice^[3]
Dosage:	200 or 800 mg/kg
Administration:	Oral gavage; 200 or 800 mg/kg; once daily; 3 months
Result:	Reduced MMP-9 activity in a dose-dependent manner.

Clinical Trial

Molecular Weight

512.94

Formula

C₂₃H₂₉ClN₂O₉

CAS No.

24390-14-5

Appearance

Solid

Color

White to yellow

SMILES

O=C(C(C1=O)=C(O)[C@@H](N(C)C)[C@]2(06)[C@@H](O)[C@]3(06)[C@@H](C)C4=C(C(C3=C(O)[C@@]21O)=O)C(O)=CC=C4)N.Cl.[0.5H2O].[0.5C2H6O]

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 1 year; -20°C, 6 months (sealed storage, away from moisture and light)

Solvent & Solubility

In Vitro:

DMSO : 240 mg/mL (467.89 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H₂O : 125 mg/mL (243.69 mM; Need ultrasonic)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	1.9495 mL	9.7477 mL	19.4955 mL
5 mM	0.3899 mL	1.9495 mL	3.8991 mL
10 mM	0.1950 mL	0.9748 mL	1.9495 mL

View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 1 year; -20°C, 6 months (sealed storage, away from moisture and light). When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 5% DMSO 40% PEG300 5% Tween-80 50% Saline
Solubility: ≥ 3 mg/mL (5.85 mM); Clear solution
Protocol 2

Add each solvent one by one: 5% DMSO 95% (20% SBE-β-CD in Saline)
Solubility: ≥ 3 mg/mL (5.85 mM); Clear solution

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: PBS
Solubility: 20 mg/mL (38.99 mM); Clear solution; Need ultrasonic

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Calculation results:

Working solution concentration: mg/mL

This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only.If necessary, please contact MedChemExpress (MCE).

Purity & Documentation

Purity: 99.19%

Data Sheet (295 KB) SDS (393 KB)

COA (262 KB) LCMS (131 KB)

Handling Instructions (2659 KB)

References

[1]. Eusebio Manchado, et al. A combinatorial strategy for treating KRAS-mutant lung cancer. Nature. 2016 Jun 30;534(7609):647-51. [Content Brief]

[2]. Anna-Luisa Luger, et al. Doxycycline Impairs Mitochondrial Function and Protects Human Glioma Cells from Hypoxia-Induced Cell Death: Implications of Using Tet-Inducible Systems. Int J Mol Sci. 2018 May 17;19(5):1504. [Content Brief]

[3]. Wilfried Briest, et al. Doxycycline ameliorates the susceptibility to aortic lesions in a mouse model for the vascular type of Ehlers-Danlos syndrome. J Pharmacol Exp Ther. 2011 Jun;337(3):621-7. [Content Brief]

[4]. Ethan Ahler, et al. Doxycycline alters metabolism and proliferation of human cell lines. PLoS One. 2013 May 31;8(5):e64561. [Content Brief]

[5]. Le Zhang, et al. Doxycycline inhibits the cancer stem cell phenotype and epithelial-to-mesenchymal transition in breast cancer. Cell Cycle. 2017 Apr 18;16(8):737-745. [Content Brief]

[6]. Manfredsson FP, et al. Tight Long-term dynamic doxycycline responsive nigrostriatal GDNF using a single rAAV vector. Mol Ther. 2009 Nov;17(11):1857-67. [Content Brief]

[7]. Kistner A, et al. Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice. Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10933-8. [Content Brief]

[8]. Niv Y. Doxycycline in Eradication Therapy of Helicobacter pylori--a Systematic Review and Meta-Analysis. Digestion. 2016;93(2):167-73. [Content Brief]

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