General:** For in vivo experiments, mice received oral (po) or intraperitoneal (ip) treatment with kaurenoic acid or vehicle (2% DMSO in saline) 30 minutes before the inflammatory stimulus, unless otherwise specified. [1]
- **Writhing Test (Acetic Acid/PBQ):** Mice were treated with kaurenoic acid (3-30 mg/kg, ip or po) 30 minutes before ip injection of acetic acid (0.8%) or PBQ (1890 μg/kg). The cumulative number of writhes was evaluated for 20 minutes. Indomethacin (5 mg/kg, ip, 40 minutes before stimulus) was used as a control. [1]
- **Formalin Test:** Mice were treated with kaurenoic acid (10 mg/kg, po) 30 minutes before intraplantar (ipl) injection of formalin (25 μL of 1.5%). The number of paw flinches and time spent licking were evaluated for 30 minutes, divided into first (0-15 min) and second (20-30 min) phases. [1]
- **Mechanical Hyperalgesia (Electronic Pressure-Meter Test):** Mice were placed in acrylic cages with wire grid floors. A hand-held force transducer with a polypropylene tip was applied perpendicularly to the central area of the hindpaw with increasing pressure. The endpoint was paw withdrawal with flinching, and the pressure intensity was recorded. The result was expressed as the delta (Δ) withdrawal threshold (in g), calculated by subtracting baseline measurements from post-stimulus measurements. [1]
- **Carrageenin-induced:** Mice were treated with kaurenoic acid (1, 3, 10 mg/kg, po) 30 minutes before ipl injection of carrageenin (100 μg/paw). Mechanical hyperalgesia was evaluated 1-5 hours after stimulus. [1]
- **CFA-induced:** Mice were treated daily with kaurenoic acid (10 mg/kg, po) starting 1 hour after ipl CFA (10 μL/paw) injection for 7 days. Mechanical hyperalgesia was evaluated daily. [1]
- **PGE2-induced:** Mice were treated with kaurenoic acid (10 mg/kg, po) 30 minutes before ipl injection of PGE2 (100 ng/paw). Mechanical hyperalgesia was evaluated 3 hours after stimulus. [1]
- **Mechanism Studies:** Mice were pretreated with L-NAME (10-90 mg/kg, ip) 60 minutes before, or with ODQ (0.3 mg/kg, ip), KT5823 (0.5 μg/mouse, ip), or glybenclamide (0.3 mg/kg, ip) before kaurenoic acid (10 mg/kg, po). After 30 minutes, carrageenin was injected, and hyperalgesia was measured. L-NAME, ODQ, and glybenclamide were diluted in 2% DMSO in saline, and KT5823 in 2% DMSO in saline. Glybenclamide was diluted in 5% Tween 80 in saline. [1]
- **Cytokine Measurement:** Mice were treated with kaurenoic acid (10 mg/kg, po) 30 minutes before carrageenin (100 μg/paw). Three hours later, paw skin tissue was collected and homogenized. TNF-α and IL-1β levels were determined by ELISA. Results expressed as pg of cytokine/paw. [1]
- **Hepatotoxicity Assessment:** Mice received daily oral treatment with vehicle (2% DMSO in saline or Tris/HCl buffer, pH 8.0), kaurenoic acid (10 mg/kg), or indomethacin (2.5 mg/kg) for 7 days. Plasma levels of AST and ALT were measured. [1]
- **Gastric MPO Activity Assessment:** Mice received daily oral treatment with vehicle, kaurenoic acid (10 mg/kg), or indomethacin (2.5 mg/kg) for 7 days. Stomach samples were collected, homogenized, and MPO activity was determined spectrophotometrically. Results expressed as number of neutrophils × 10^7 / mg of tissue. [1]
- **Motor Performance (Rota-rod Test):** Mice were selected 24 hours prior based on their ability to stay on a rotating rod (22 rpm) for 120 s. Treated animals (kaurenoic acid, 10 mg/kg, po) were tested at 1.5, 3.5, and 5.5 hours post-treatment, with a cutoff time of 120 s. [1]
- **Hot Plate Test:** Mice were placed on a hot plate maintained at 55°C. The reaction time (jumping or paw licking) was recorded before and 1.5, 3.5, and 5.5 hours after treatment (kaurenoic acid, 10 mg/kg, po), with a cutoff of 30 s to avoid tissue damage. Morphine (8 mg/kg, ip) was used as a positive control. [1]

[1]. Kaurenoic acid from Sphagneticola trilobata Inhibits Inflammatory Pain: effect on cytokine production and activation of the NO-cyclic GMP-protein kinase G-ATP-sensitive potassium channel signaling pathway. J Nat Prod. 2012 May 25;75(5):896-904.

Ent-kaur-16-en-19-oic acid is an ent-kaurane diterpenoid compound, namely ent-kauran-19-oic acid, which has a double bond at position 16(17); it possesses anticancer and anti-HIV-1 activities. It can be used as an anti-HIV-1 drug, an antitumor drug, and a plant metabolite. It is the conjugate acid of ent-kaur-16-en-19-oate. Kaurenoic acid has been reported to exist in Disynaphia multicrenulata, Eleutherococcus koreanus, and several other organisms with relevant data.

C20H30O2

302.46

302.224

6730-83-2

73062

White to off-white solid powder

1.1±0.1 g/cm3

426.6±34.0 °C at 760 mmHg

202.8±20.3 °C

0.0±2.2 mmHg at 25°C

1.550

6.37

1

2

1

22

538

6

C[C@@]12CCC[C@@]([C@H]1CC[C@]34[C@H]2CC[C@H](C3)C(=C)C4)(C)C(=O)O

NIKHGUQULKYIGE-OTCXFQBHSA-N

InChI=1S/C20H30O2/c1-13-11-20-10-7-15-18(2,16(20)6-5-14(13)12-20)8-4-9-19(15,3)17(21)22/h14-16H,1,4-12H2,2-3H3,(H,21,22)/t14-,15+,16+,18-,19-,20-/m1/s1

(1S,4S,5R,9S,10R,13R)-5,9-dimethyl-14-methylidenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylic acid

CCRIS 1514 Kauren-19-oic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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

DMSO : ~100 mg/mL (~330.63 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (8.27 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 25.0 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 2: ≥ 2.5 mg/mL (8.27 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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