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CTPI-2

Cat No.:V9925 Purity: ≥98%
CTPI-2 is a novel, potent, specific, andthird-generation mitochondrial citrate carrier SLC25A1 inhibitor with a KD of 3.5 μM.
CTPI-2
CTPI-2 Chemical Structure CAS No.: 68003-38-3
Product category: Mitochondrial Metabolism
This product is for research use only, not for human use. We do not sell to patients.
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1g
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Product Description

CTPI-2 (CTPI2) is a novel, potent, specific, and third-generation mitochondrial citrate carrier SLC25A1 inhibitor with a KD of 3.5 μM. CTPI-2 inhibits glycolysis, PPARγ, and its downstream target the glucose transporter GLUT4. CTPI-2 halts salient alterations of NASH reverting steatosis, preventing the evolution to steatohepatitis, reducing inflammatory macrophage infiltration in the liver and adipose tissue, and starkly mitigating obesity induced by a high-fat diet.

Biological Activity I Assay Protocols (From Reference)
ln Vivo
The specific glycolysis regulator CTPI-2 restricts the metabolic adaptability of cancer stem cells (CSCs). Non-small cell lung cancer (NSCLC) in vivo model tumor growth is inhibited by intraperitoneal administration of CTPI-2 (26 mg/kg) [1]. ?In a prevention study, CTPI-2 (50 mg/kg; intraperitoneal injection; every other day for 12 weeks) entirely avoided weight gain, and in a reversal study, it significantly reduced weight [2]. ?CTPI-2 restores normal glucose tolerance and avoids steatohepatitis. In addition to boosting anti-inflammatory IL-4 and IL-10, CTPI-2 decreases circulating levels of IL-6 and decreases interferon gamma-induced monocyte chemoattractant protein 1 and monokine that draw neutrophils and monocytes. Citrate depots, adipogenesis, and gluconeogenesis pathways are all regulated by CTPI-2 [2].
Animal Protocol
Animal/Disease Models: C57BL/6J mice (HFD-fed mice) [2]
Doses: 50 mg/kg
Route of Administration: Via intraperitonealroute on alternate days for 12 weeks
Experimental Results: Complete avoidance of weight gain in prevention study, and resulted in significant weight loss regression studies.
References

[1]. Inhibition of the mitochondrial citrate carrier, Slc25a1, reverts steatosis, glucose intolerance, and inflammation in preclinical models of NAFLD/NASH. Cell Death Differ. 2020;27(7):2143-2157.

[2]. The mitochondrial citrate carrier, SLC25A1, drives stemness and therapy resistance in non-small cell lung cancer. Cell Death Differ. 2018;25(7):1239-1258.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C13H11N2O6SCL
Molecular Weight
358.75424
Exact Mass
355.987
CAS #
68003-38-3
Related CAS #
68003-38-3;
PubChem CID
106350
Appearance
Off-white to light yellow solid powder
Density
1.66g/cm3
Boiling Point
564.8ºC at 760 mmHg
Flash Point
295.4ºC
Index of Refraction
1.678
LogP
4.424
Hydrogen Bond Donor Count
2
Hydrogen Bond Acceptor Count
7
Rotatable Bond Count
4
Heavy Atom Count
23
Complexity
558
Defined Atom Stereocenter Count
0
InChi Key
NJTHPOSQGFJTDP-UHFFFAOYSA-N
InChi Code
InChI=1S/C13H9ClN2O6S/c14-10-6-5-8(7-12(10)16(19)20)23(21,22)15-11-4-2-1-3-9(11)13(17)18/h1-7,15H,(H,17,18)
Chemical Name
2-[(4-chloro-3-nitrophenyl)sulfonylamino]benzoic acid
HS Tariff Code
2934.99.9001
Storage

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
DMSO : ~125 mg/mL (~350.40 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.83 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 2: ≥ 2.08 mg/mL (5.83 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C,1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (5.83 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.


 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 2.7875 mL 13.9373 mL 27.8746 mL
5 mM 0.5575 mL 2.7875 mL 5.5749 mL
10 mM 0.2787 mL 1.3937 mL 2.7875 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

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What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
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What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
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Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

Biological Data
  • Slc25a1 inhibition with CTPI-2 ameliorates steatosis and liver injury. a Time-course experiments showing the livers of HFD-fed mice treated with vehicle (top panels) or with CTPI-2 (bottom panels). The weeks of diet exposure are indicated at the top; the times of treatment with CTPI-2 are at the bottom. Time 0 shows the liver histology when CTPI-2 treatment was initiated, after 12 weeks of HFD. The last panel on the right shows a normal liver derived from CD-fed mice. Rectangles show enlarged fields and arrows point to ballooning hepatocytes, when detected. b, c Total serum cholesterol levels and ALT levels measured with the Heska Element DC blood chemistry analyzer. d Serum levels of triglycerides measured with LC-MS. e Quantification of steatosis from the time-course experiments. The percentage of steatosis in CTPI-2 treated mice is shown on the top of each bar graph. f, g Quantification of liver steatosis indicated as % per field (f) or steatosis grade (g). Quantification was performed on 3–5 mice per group and on at least 2–3 fields per mouse. Steatosis grade was assessed as: grade 0: <5%; grade 1: 5–33%; grade 2: 33–50%; and grade 3: >50%. h Representative images of HFD-fed mice (left panel) and of three different mice treated with CTPI-2. Rectangles show enlarged fields of the images and arrows indicate hepatocyte ballooning, when present. *p ≤ 0.05, **p ≤ 0.01, ***p ≤0 0.001.[1].Tan M, et al. Inhibition of the mitochondrial citrate carrier, Slc25a1, reverts steatosis, glucose intolerance, and inflammation in preclinical models of NAFLD/NASH. Cell Death Differ. 2020;27(7):2143-2157.
  • CTPI-2 normalizes glucose tolerance and insulin sensitivity. a Fasting glucose levels in control diet (C, white bars), HFD+Vehicle (V, grey bars), or HFD+CTPI-2 (T, red bars) mice (n = 3–6). b Insulin levels (in μg/ml) of animals treated with CTPI-2 for approximately 11 weeks (n = 2–3). c Glucose tolerance test performed at the indicated time points of CTPI-2 treatment (n = 3–4). Grey lines indicate control diet; black lines indicate HFD+vehicle; and red lines indicate HFD+CTPI-2. d Insulin tolerance test in the indicated mice groups. e Expression levels of Slc25a1 (shown at two different exposure times, Exp.1 and Exp.2) in the indicated organs and treatment conditions. f Expression levels of Slc25a1 in the visceral adipose tissue of mice fed with chow (C), high-glucose (HG), or low-glucose (LG) diets (see “Materials and methods” for diet composition). g Fasting glucose levels in mice fed as indicated (n = 4–6). *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.[1].Tan M, et al. Inhibition of the mitochondrial citrate carrier, Slc25a1, reverts steatosis, glucose intolerance, and inflammation in preclinical models of NAFLD/NASH. Cell Death Differ. 2020;27(7):2143-2157.
  • CTPI-2 influences inflammatory pathways. a Serum levels of the indicated interleukins and chemoattractant factors from mice fed the HFD and receiving vehicle (V) or CTPI-2 (T). b Quantification of liver macrophages in mice fed with control diet (C), or with the HFD and treated with vehicle (V) or CTPI-2 (T). Quantification was performed on 2–3 mice per group and from multiple fields per mouse with the ImageJ program. c Representative IHC images of F4/80 staining in the livers of the indicated treatment groups. d, e Quantification of macrophages in the WAT (d) and representative IHC images of H&E staining in the visceral adipose tissue, with crown structures indicated by arrows (e). f, g mRNA levels of the indicated genes in the livers of vehicle (black) or CTPI-2 treated mice (red). h Representative images of Picro-Sirius Red staining in the liver of the indicated mice. iNOS inducible nitric oxide synthase, TNFα tumor necrosis factor alpha, IFNγ interferon gamma, Interleukins 10–13, MRC1 Macrophage Mannose Receptor 1, FN1 fibronectin 1, Arg1 Arginase 1, Col4 Collagen 4, Col1a Collagen 1a, KRT19 Keratin 19, PDGFR platelet-derived growth factor receptor alpha, CDH1 cadherin-1. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.[1].Tan M, et al. Inhibition of the mitochondrial citrate carrier, Slc25a1, reverts steatosis, glucose intolerance, and inflammation in preclinical models of NAFLD/NASH. Cell Death Differ. 2020;27(7):2143-2157.
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