2A3 (Synonyms: 2-aminopyridine-3-carboxylic acid imidazolide)

Cat. No.: HY-164899 Purity: 97.0%: Data Sheet
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2A3 (2-aminopyridine-3-carboxylic acid imidazolide) is a T cell activator that specifically binds to CEACAM6 and CEACAM5. 2A3 exhibits enzymatic activity that catalyzes the glucuronidation of specific substrates (e.g., 1-naphthol), and possesses significant cytotoxic activity. When integrated into CAR T cells or used alone, 2A3 acts by inducing cytokine release, degranulation, and direct cytotoxicity. 2A3 kills pancreatic and breast cancer cells with high target antigen expression in vitro, and significantly inhibits the growth of pancreatic cancer xenografts in vivo. 2A3 broadly targets malignant tumors with overexpressed CEACAM5, CEACAM6, or co-expressed both, and shows high expression mainly in tissues such as the liver and colon. 2A3 serves as an important research tool for the immunotherapy of pancreatic and breast cancer. 2A3 is a novel SHAPE reagent, which can be used for the analysis of RNA structure both in vitro and in vivo. 2A3 is an electrophilic chemical probe that acylates the 2'-OH in the RNA backbone. 2A3 can be used for RNA SHAPE-MaP experiments and is capable of analyzing the RNA secondary structures at single nucleotide resolution.

For research use only. We do not sell to patients.

2A3 Chemical Structure

CAS No. : 2765091-45-8

Size	Stock	Quantity

Free Sample (0.1 - 0.2 mg)	Apply Now
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO ready for reconstitution	In-stock	Estimated Time of Arrival: December 31
Solution
10 mM * 1 mL in DMSO	In-stock	Estimated Time of Arrival: December 31
Solid
5 mg	In-stock	Estimated Time of Arrival: December 31
10 mg	In-stock	Estimated Time of Arrival: December 31
25 mg	In-stock	Estimated Time of Arrival: December 31
50 mg	In-stock	Estimated Time of Arrival: December 31
100 mg	In-stock	Estimated Time of Arrival: December 31
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Description

2A3 (2-aminopyridine-3-carboxylic acid imidazolide) is a T cell activator that specifically binds to CEACAM6 and CEACAM5. 2A3 exhibits enzymatic activity that catalyzes the glucuronidation of specific substrates (e.g., 1-naphthol), and possesses significant cytotoxic activity. When integrated into CAR T cells or used alone, 2A3 acts by inducing cytokine release, degranulation, and direct cytotoxicity. 2A3 kills pancreatic and breast cancer cells with high target antigen expression in vitro, and significantly inhibits the growth of pancreatic cancer xenografts in vivo. 2A3 broadly targets malignant tumors with overexpressed CEACAM5, CEACAM6, or co-expressed both, and shows high expression mainly in tissues such as the liver and colon. 2A3 serves as an important research tool for the immunotherapy of pancreatic and breast cancer^[1]^[2]. 2A3 is a novel SHAPE reagent, which can be used for the analysis of RNA structure both in vitro and in vivo^[3]. 2A3 is an electrophilic chemical probe that acylates the 2'-OH in the RNA backbone. 2A3 can be used for RNA SHAPE-MaP experiments and is capable of analyzing the RNA secondary structures at single nucleotide resolution.

In Vitro

2A3-CAR lentivirus (1 virion per cell; 24 h, 48 h post-T-cell activation) results in consistent expression of the 2A3-CAR on ~90% of the activated human T-cell population^[1].
2A3-CAR T-cells (effector-to-target ratio 3:1, 5:1; 24 h co-culture) mediate potent cytotoxicity, IFN-γ secretion, and degranulation against CEACAM5/6-positive human pancreatic (BxPC-3, Capan-1, MIA PaCa-2) and breast (MCF7, SK-BR-3) cancer cell lines, but not CEACAM5/6-negative MDA-MB-231 cells^[1].
2A3-CAR T-cells (effector-to-target ratio 5:1; 24 h co-culture) retain cytotoxic activity against CEACAM6-knockout MCF7 breast cancer cells, indicating CEACAM6 is not the sole target of the 2A3-CAR^[1].
2A3-CAR T-cells (effector-to-target ratio 10:1; 12 h, 24 h co-culture) mediate potent cytotoxicity against MDA-MB-231 cells overexpressing either CEACAM5 or CEACAM6, with stronger activity against CEACAM6-overexpressing cells^[1].

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

Cell Cytotoxicity Assay^[1]

Cell Line:	Human pancreatic cell lines (BxPC-3, Capan-1, MIA PaCa-2), human breast cancer cell lines (MCF7, SK-BR-3, MDA-MB-231)
Concentration:	Effector-to-target ratio 3:1 (pancreatic cell lines); effector-to-target ratio 5:1 (breast cancer cell lines)
Incubation Time:	24 h (co-culture)
Result:	Exerted potent cytotoxic effects on BxPC-3, Capan-1, MIA PaCa-2, MCF7, and SK-BR-3 cell lines, but not MDA-MB-231. Increased IFN-γ concentrations significantly in supernatants of BxPC-3, Capan-1, MIA PaCa-2, MCF7, and SK-BR-3 cells co-cultured with 2A3-CAR T-cells, but not MDA-MB-231. Showed a trend toward increased CD107a expression after co-culture with BxPC-3, Capan-1, MIA PaCa-2, MCF7, and SK-BR-3 cells, but not MDA-MB-231.

Cell Cytotoxicity Assay^[1]

Cell Line:	MDA-MB-231 human breast cancer cell line derivatives overexpressing CEACAM5 or CEACAM6
Concentration:	Effector-to-target ratio 10:1
Incubation Time:	12 h, 24 h (co-culture)
Result:	Mediated significant cytotoxic effects against both CEACAM5-overexpressing and CEACAM6-overexpressing MDA-MB-231 cells, compared to mock or untransduced T-cells. Showed cytotoxic effect against CEACAM6-overexpressing MDA-MB-231 cells that was approximately six times stronger than against CEACAM5-overexpressing cells at 12 h and 24 h post-co-culture. Showed no significant cytotoxicity against unmodified MDA-MB-231 cells.

In Vivo

2A3-CAR T-cells (10⁷ cells/mouse; i.v.; days 1, 8, 15 (early); days 12, 20, 26 (late)) exert high in vivo antitumor efficacy against BxPC-3 pancreatic cancer xenografts in both early and late intervention models, inducing significant tumor growth inhibition and complete regression in some mice^[1].

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

Animal Model:	CIEA NOG mice with Pancreatic cancer (female, 5 weeks of age)^[1]
Dosage:	10⁷ cells/mouse (early intervention model); 10⁷ cells/mouse (late intervention model)
Administration:	i.v.; days 1, 8, 15 (early intervention model); days 12, 20, 26 (late intervention model)
Result:	Significantly decreased BxPC-3 xenograft growth (p < 0.0003 vs control T-cells on day 30), with complete tumor regression observed in one mouse (early intervention model). Controlled or regressed tumor growth (p < 0.02 vs control T-cells on day 34), with complete tumor regression observed in two mice (late intervention model). Caused no generalized toxicities based on body weight measurements.

Molecular Weight

188.19

Formula

C₉H₈N₄O

CAS No.

2765091-45-8

Appearance

Solid

Color

White to off-white

SMILES

O=C(C1=CC=CN=C1N)N2C=NC=C2

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, protect from light, stored under nitrogen

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

Solvent & Solubility

In Vitro:

DMSO : 20 mg/mL (106.28 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	5.3138 mL	26.5689 mL	53.1378 mL
5 mM	1.0628 mL	5.3138 mL	10.6276 mL
10 mM	0.5314 mL	2.6569 mL	5.3138 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, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Molarity Calculator
Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

Concentration _(start)

Volume _(start)

Concentration _(final)

Volume _(final)

In Vivo:

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: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2 mg/mL (10.63 mM); Clear solution

This protocol yields a clear solution of ≥ 2 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Protocol 2

Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 2 mg/mL (10.63 mM); Clear solution

This protocol yields a clear solution of ≥ 2 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.0 mg/mL) to 900 μL Corn oil, and mix evenly.

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.

Please enter your animal formula composition:

DMSO +

Tween-80 +

Saline

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Calculation results:

Working solution concentration: mg/mL

Method for preparing stock solution: mg drug dissolved in μL DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

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).

Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.

If the continuous dosing period exceeds half a month, please choose this protocol carefully.

Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.

Purity & Documentation

Purity: 97.0%

Select Batch:

Data Sheet (283 KB) SDS (252 KB)

COA (253 KB) HNMR (241 KB) MS (70 KB)

Handling Instructions (2659 KB)

References

[1]. Jancewicz I, Śmiech M, Winiarska M, et al. New CEACAM-targeting 2A3 single-domain antibody-based chimeric antigen receptor T-cells produce anticancer effects in vitro and in vivo[J]. Cancer immunology, immunotherapy, 2024, 73(2): 30.

[2]. Bushey R T, Dluzen D F, Lazarus P. Importance of UDP-glucuronosyltransferases 2A2 and 2A3 in tobacco carcinogen metabolism[J]. Drug Metabolism and Disposition, 2013, 41(1): 170-179.

[3]. Marinus T, Fessler AB, Ogle CA, Incarnato D. A novel SHAPE reagent enables the analysis of RNA structure in living cells with unprecedented accuracy. Nucleic Acids Res. 2021;49(6):e34. [Content Brief]

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

DMSO	1 mM	5.3138 mL	26.5689 mL	53.1378 mL	132.8445 mL
	5 mM	1.0628 mL	5.3138 mL	10.6276 mL	26.5689 mL
	10 mM	0.5314 mL	2.6569 mL	5.3138 mL	13.2844 mL
	15 mM	0.3543 mL	1.7713 mL	3.5425 mL	8.8563 mL
	20 mM	0.2657 mL	1.3284 mL	2.6569 mL	6.6422 mL
	25 mM	0.2126 mL	1.0628 mL	2.1255 mL	5.3138 mL
	30 mM	0.1771 mL	0.8856 mL	1.7713 mL	4.4281 mL
	40 mM	0.1328 mL	0.6642 mL	1.3284 mL	3.3211 mL
	50 mM	0.1063 mL	0.5314 mL	1.0628 mL	2.6569 mL
	60 mM	0.0886 mL	0.4428 mL	0.8856 mL	2.2141 mL
	80 mM	0.0664 mL	0.3321 mL	0.6642 mL	1.6606 mL
	100 mM	0.0531 mL	0.2657 mL	0.5314 mL	1.3284 mL

2A3 Related Classifications

Help & FAQs

Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

2A32765091-45-82-aminopyridine-3-carboxylic acid imidazolideTransmembrane GlycoproteinCEACAM5CEACAM6pancreatic cancerT-cellsbreast cancerMIA PaCa-2MCF7BxPC-3NOG miceCapan-1Inhibitorinhibitorinhibit

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2A3 (Synonyms: 2-aminopyridine-3-carboxylic acid imidazolide)

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