TACE (ADAM17); MMP
TAPI-1 is an inhibitor of ADAM17 (TACE). No IC50, Ki, EC50, or DC50 values are reported in the provided literature. [3][4]

TAPI-1 inhibits the release of soluble forms of TNF-alpha, p60 TNFR, and IL-6R from human peripheral blood monocytes and the monocytic cell line THP-1, both in response to stimulation and PMA. Additionally, TAPI prevents monocytes from shedding TNF-alpha and p60 TNFR in response to LPS.[1]
TAPI-1 prevents co-transfected APP from constitutively releasing in a TACE-dependent manner (695).[2]
TAPI-1 reduces the effects of Ang II-induced EGFR transactivation and cell division in human HSC line LI90.[3]
TAPI-1 in combination with the EGFR inhibitor AG1478 shows a deactivated AREG/EGFR/ERK signaling pathway and decreases the release of pro-inflammatory cytokines in salivary gland-derived epithelial cells from pSS.[4]

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In human hepatic stellate LI90 cells, pre-incubation with TAPI-1 (20 μM, 45 min) suppressed angiotensin II (10⁻⁶ M)-induced phosphorylation of EGFR (at 5 min) as determined by Western blotting using an anti-phospho-EGFR (pTyr1068) antibody. [3]
- In the same LI90 cells, TAPI-1 (20 μM, 45 min pre-incubation) significantly attenuated angiotensin II (10⁻⁶ M, 60 h)-induced cell proliferation measured by the CellTiter-Glo Luminescent Cell Viability assay. [3]
- In primary rat hepatic stellate cells, TAPI-1 (20 μM, 45 min pre-incubation) inhibited angiotensin II (10⁻⁶ M)-induced EGFR phosphorylation (at 5 min) shown by Western blotting. [3]
- In primary rat hepatic stellate cells, TAPI-1 (20 μM, 45 min pre-incubation) repressed angiotensin II (10⁻⁶ M, 60 h)-induced cell proliferation measured by an ATP-based viability assay. [3]
- In human salivary gland epithelial cells (SGEC) derived from primary Sjögren’s syndrome (pSS) patients, treatment with TAPI-1 (10 μM, 24 h) significantly reduced the phosphorylation levels of ERK1/2 as demonstrated by Western blotting using an anti-phospho-ERK1/2 antibody. [4]
- In pSS SGEC, TAPI-1 (10 μM, 24 h) significantly decreased the secretion of multiple pro-inflammatory cytokines (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17A, IFNγ, TNF-α, GM-CSF) into the cell culture supernatant, as measured by a human cytokine Multi-Analyze ELISA Array kit. [4]

TAPI-1 is an ADAM17/TACE inhibitor that prevents cytokine receptors from being shed.

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The inhibition of the pressor response to big ET-1 (1 nmol/kg) by Phosphoramidon in ganglion-blocked anesthetized rats was dose-dependent, with an ID50 of approximately 5 mg/kg. At 1 mg/kg, the pressor response was reduced to ~90% of control; at 10 mg/kg, to ~30% of control; and at 30 mg/kg, to ~0% of control [5].

For HPLC analysis, partially purified ECE from rabbit lung (480 µg protein) was incubated with 3.75 µM big ET-1-(1-39) and 5 µM thiorphan in 1.0 mL of 50 mM Tris-HCl (pH 7.5) at 37°C. A parallel reaction was run containing 100 µM Phosphoramidon. At 1-hour intervals, 100 µL aliquots were removed and immediately analyzed by reverse-phase HPLC to monitor the conversion of big ET-1 (retention time 34.3 min) to ET-1 (retention time 36.8 min) [5].

Each well of 96-well plates contains 5,000 seeded cells, and the CellTiter-Glo Luminescent Cell Viability Assay is used to determine how viable the cells are. After 24 hours of serum deprivation, the cells undergo Ang II treatment for 60 hours, both with and without prior inhibitor and antagonist pretreatment. Next, each well on the plate is filled with the assay substrates, and a luminometer is used to assess the samples.

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LI90 human activated hepatic stellate cells: Cells were seeded at 5,000 cells per well in 96-well plates. After 24 h of serum deprivation, cells were pre-incubated with TAPI-1 at 20 μM for 45 min, then treated with angiotensin II at 10⁻⁶ M for 60 h. Cell viability/proliferation was assessed using the CellTiter-Glo Luminescent Cell Viability assay, and luminescence was measured with a luminometer. For Western blotting, cells were treated similarly (45 min pre-incubation with 20 μM TAPI-1 followed by 5 min stimulation with 10⁻⁶ M angiotensin II), then lysed in ice-cold buffer. Lysates were subjected to SDS-PAGE and transferred to PVDF membranes, which were probed with anti-phospho-EGFR (pTyr1068) and anti-EGFR antibodies. [3]
- Primary rat hepatic stellate cells: Isolated from normal Wistar rats and used at passages 4-6 (in vitro activation). Cells were serum-starved for 24 h, then pre-incubated with TAPI-1 at 20 μM for 45 min, followed by angiotensin II (10⁻⁶ M) stimulation for 5 min (for Western blot) or 60 h (for proliferation assay). Western blotting for phospho-EGFR and total EGFR was performed as described for LI90 cells. Proliferation was measured using an ATP-based luminescent cell viability assay. [3]
- Human salivary gland epithelial cells (pSS SGEC): Epithelial cells were cultured from labial minor salivary gland biopsies of primary Sjögren’s syndrome patients. Cells were treated with TAPI-1 at 10 μM for 24 h. After treatment, cell lysates were collected for Western blot analysis using antibodies against phospho-ERK1/2 and total ERK1/2. Separately, cell culture supernatants were collected for cytokine quantification using a human cytokine Multi-Analyze ELISA Array kit that measured IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17A, IFNγ, TNF-α, and GM-CSF. Absorbance was read at 450 nm. [4]

Male Sprague-Dawley rats
1 μg
i.c.v.

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In another setup, big ET-1 at 1 nmol/kg or ET-1 at 1 nmol/kg was injected intravenously into ganglion-blocked anesthetized rats. Phosphoramidon at 30 mg/kg was injected just prior to the administration of the peptide to assess its effect on the pressor response [5].

[1]. J Immunol . 1995 Dec 1;155(11):5198-205.

[2]. Biochem J . 2001 Aug 1;357(Pt 3):787-94.

[3]. Life Sci . 2014 Mar 3;97(2):137-44.

[4]. Clin Exp Med . 2015 May;15(2):215-25.

[5]. Immunobiology . 2020 Mar;225(2):151887.

[6]. Hypertension . 2019 Jul;74(1):63-72.

TAPI-1 is identified as a TNF-α converting enzyme (TACE)/ADAM17 inhibitor. It is used to demonstrate that ADAM17 mediates the shedding of EGFR ligands (particularly amphiregulin) in response to angiotensin II stimulation in hepatic stellate cells, thereby enabling EGFR transactivation and subsequent cell proliferation. [3]
- In primary Sjögren’s syndrome, TAPI-1 is used to show that ADAM17 activity drives the amphiregulin/EGFR/ERK signaling pathway, leading to the production of pro-inflammatory cytokines by salivary gland epithelial cells. Blockade of ADAM17 with TAPI-1 reduces this inflammatory epithelial response. [4]
- The literature suggests that ADAM17 could be a promising therapeutic target in liver fibrosis and autoimmune epithelitis, with TAPI-1 serving as a tool compound to validate this concept. [3][4]

C26H37N5O5

499.602486371994

499.279

171235-71-5

TAPI-1;163847-77-6

9827273

White to off-white solid

1.2±0.1 g/cm3

1.581

1.29

6

6

13

36

746

2

C[C@@H](C(=O)NCCN)NC(=O)[C@H](CC1=CC2=CC=CC=C2C=C1)NC(=O)C(CC(C)C)CC(=O)NO

AWNBSWDIOCXWJW-OWHMDLSXSA-N

InChI=1S/C26H37N5O5/c1-16(2)12-21(15-23(32)31-36)25(34)30-22(26(35)29-17(3)24(33)28-11-10-27)14-18-8-9-19-6-4-5-7-20(19)13-18/h4-9,13,16-17,21-22,36H,10-12,14-15,27H2,1-3H3,(H,28,33)(H,29,35)(H,30,34)(H,31,32)/t17-,21?,22-/m0/s1

N-[(2S)-1-[[(2S)-1-(2-aminoethylamino)-1-oxopropan-2-yl]amino]-3-naphthalen-2-yl-1-oxopropan-2-yl]-N'-hydroxy-2-(2-methylpropyl)butanediamide

TAPI-1; TAPI 1; TAPI;163847-77-6

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)