Wilforlide A (30-150 μg/kg) significantly inhibits tampon-induced granulation formation in rats [1]. Wilforlide A (60-300 μg/kg) significantly inhibits xylene-induced ear swelling in ICR mice. The inhibition rates of Wilforlide A at high and low doses were 50.9% and 12.29% respectively [1].
High-dose Wilforlide A (300 μg/kg) significantly inhibited xylene-induced mice ear swelling with an edema rate of 1.28 ± 0.28 mg and inhibition rate of 50.90% (P < 0.05 compared to control). Low-dose (60 μg/kg) showed no significant difference. [1]
High-dose Wilforlide A (150 μg/kg) significantly inhibited tampon-induced rat granulation with granulation weight of 47.68 ± 11.53 mg (P < 0.05 vs control). Low-dose (30 μg/kg) also showed inhibition (51.15 ± 9.04 mg, P < 0.05 vs control). [1]
In carrageenan-induced rat paw edema, all experimental groups reached edema peak at 4 h with obvious inhibition at 6 h, but none of the groups showed significant difference from control. [1]
For immune suppression: Wilforlide A (high-dose 300 μg/kg, low-dose 60 μg/kg) did not significantly reduce mouse hemolysin response (HC50: high-dose 115.51, low-dose 124.18 vs control 149.85; no significant difference from control group) nor carbon elimination phagotrophy index (high-dose 4.38 ± 0.53, low-dose 4.21 ± 0.87 vs control 4.45 ± 0.53; not significant). In contrast, triptolide (TP) and tripterygium glycosides (TG) showed significant immune suppression. Wilforlide A also did not significantly affect delayed-type hypersensitivity or immune organ weights (data not shown). [1]

For carrageenan-induced rat pedal swelling: Male SD rats (body weight 160 ± 10 g) were randomly divided into 5 groups (n=10 per group) and administered by oral gavage. The drug dosage was adjusted based on clinical equivalency dosage acting on body surface area, and the dispensing solution was 0.5% CMC-Na. The control group received saline. After 30 min, 0.1 mL of 10 g/L carrageenan was injected into the right hind foot under the plantar aponeurosis. Paw volume was measured immediately and at 1, 2, 4, and 6 h after injection using the displacement technique with a calibrated glass tube. Edema rate and inhibition rate were calculated. [1]
For xylene-induced mice ear swelling: Male ICR mice (body weight 20 ± 2 g) were administered test drugs by oral gavage. After 30 min, 30 μL xylene (15 μL on each side) was smeared on the right ear, while the left ear received no treatment. After 1 h, mice were sacrificed, both ears were cut, and the middle of the ears was perforated with a hole puncher and weighed. Edema rate and inhibition rate were calculated. [1]
For tampon-induced rat granulation: Rats were administered by oral gavage once daily for 7 days. After the first administration, a highly compressed steam-sterilized tampon was inserted into the bilateral axillary fossa. The rats were sacrificed 24 h after the last administration, the tampons were removed, baked at 40°C for 12 h, and weighed. The weight of granulation swelling was calculated as post-insertion weight minus pre-insertion weight. [1]
For hemolysin formation reaction: Mice were administered by oral gavage once daily for 7 days. After the second administration, each mouse was injected intraperitoneally with 0.2 mL sheep red blood cell (SRBC) suspension to immunize. On day 8, blood was drawn from the eyeball, centrifuged at 3000 rpm for 10 min. Serum was diluted 800-fold, then 0.5 mL SRBC suspension was added and cooled in an ice bath, followed by 1 mL complement (guinea pig serum diluted 10-fold). The mixture was incubated in a 37°C water bath for 10 min, then the reaction was terminated in an ice bath and centrifuged at 4000 rpm for 10 min. One mL of supernatant was taken, mixed with 3 mL siderosis albumen reagent, and absorbance was measured at 540 nm. HC50 was calculated. Thymus and spleen were weighed and organ indices calculated as (organ weight/body weight) × 100%. [1]
For delayed hypersensitivity immune: Mice were administered orally once daily for 7 days. After the first administration, the middle abdomen was depilated and smeared with 50 g/20 μL DNFB. On day 6, 10 g/20 μL DNFB was smeared on the right ear (10 μL each side), while the left ear was smeared with 20 μL acetone as control. After 30 h, mice were sacrificed, both ears were cut, and the middle was perforated and weighed. Edema rate and inhibition rate were calculated. [1]
For carbon elimination from plasma: Mice were administered orally once daily for 7 days. One hour after the last administration, 0.1 mL/10 g Indian ink was injected into the caudal vein. Blood (20 μL) was drawn from the eyeball at 1 min (t1) and 5 min (t2), mixed with 2 mL 0.1% Na2CO3, and absorbance (OD1, OD2) was measured at 280 nm. Mice were sacrificed, liver and spleen were weighed. Clearance index (K) and phagotrophy index (a) were calculated. [1]

[1]. Comparative study on the anti-inflammatory and immune suppressive effect of Wilforlide A. Fitoterapia.

Wilford A has been reported in Tripterygium wilfordii, Tripterygium hypoglaucum, and other organisms with available data.

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Wilforlide A (T1) is a triterpene purified compound isolated from Tripterygium wilfordii Hook f. (TWHF). It serves as a quality control standard for Tripterygium Glycosides (TG) and is officially listed in the Drug Standard of the Ministry of Public Health of the People's Republic of China (WS3-B-3350-98). This study compared its anti-inflammatory and immune suppressive effects with triptolide (TP) and TG. The results showed that Wilforlide A has obvious anti-inflammatory effect but no significant immune suppressive activity, in contrast to TP and TG. The authors suggest that using the content of Wilforlide A alone cannot evaluate the overall quality of TG, and the content of TP should also be controlled as a supplement standard to ensure safe and effective medication. TP has a narrow safety range with activity and toxicity closely related and inseparable. [1]

C30H46O3

454.6844

454.344

84104-71-2

158477

White to off-white solid powder

1.1±0.1 g/cm3

555.0±50.0 °C at 760 mmHg

318 °C

208.6±22.9 °C

0.0±3.4 mmHg at 25°C

1.563

7.47

1

3

0

33

935

10

C[C@]12CC[C@@H](C([C@@H]1CC[C@@]3([C@@H]2CC=C4[C@]3(CC[C@@]5([C@H]4C[C@]6(C[C@@H]5OC6=O)C)C)C)C)(C)C)O

HHQJBWYXBWOFJY-YLXTXNMFSA-N

InChI=1S/C30H46O3/c1-25(2)20-10-13-30(7)21(28(20,5)12-11-22(25)31)9-8-18-19-16-26(3)17-23(33-24(26)32)27(19,4)14-15-29(18,30)6/h8,19-23,31H,9-17H2,1-7H3/t19-,20-,21+,22-,23-,26-,27+,28-,29+,30+/m0/s1

(2S,5S,5aR,7aS,7bR,9aR,11S,13aR,13bR,15bS)-11-hydroxy-2,5a,7a,7b,10,10,13a-heptamethyl-1,5,5a,6,7,7a,7b,8,9,9a,10,11,12,13,13a,13b,14,15b-octadecahydro-2,5-methanochryseno[2,1-c]oxepin-3(2H)-one

Wilforlide A; Regelide; Abruslactone A;

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)

DMSO : ~1 mg/mL (~2.20 mM)

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.)