Glucocorticoid receptor (GR); The target of Hydrocortisone 17-valerate is the intracellular glucocorticoid receptor (GR). Upon binding to GR, the active component activates the expression of anti-inflammatory genes while suppressing the transcription of pro-inflammatory genes. Additionally, the drug inhibits phospholipase A2 activity and cyclooxygenase-2 (COX-2) expression, thereby blocking the synthesis of the inflammatory mediators prostaglandins and leukotrienes. By inducing the production of lipocortins, it also reduces the migration of inflammatory cells to the site of inflammation and inhibits the release of cytokines involved in the inflammatory response.

Hydrocortisone 17-valerate exhibits anti-inflammatory activity in vitro through binding to the glucocorticoid receptor. As a prodrug of hydrocortisone, its active form's binding affinity to the receptor determines its in vitro biological effects. Research indicates that this compound inhibits the production of pro-inflammatory cytokines such as tumor necrosis factor (TNF) and interleukin-1 (IL-1). In in vitro cellular models, it induces the production of phospholipase A2 inhibitory proteins (lipocortins), thereby controlling the biosynthesis of potent inflammatory mediators such as prostaglandins and leukotrienes, consequently reducing inflammatory responses.

Superior vasoconstrictor activity compared to hydrocortisone in human skin (blanching assay)[2]
Significant reduction in eczema severity scores vs. baseline (p<0.001) in atopic dermatitis patients[1][4]
Hydrocortisone valerate 0.2% ointment showed significantly better efficacy than hydrocortisone 1% in treating eczematous dermatitis (p<0.01)[2]
Equivalent efficacy to betamethasone valerate 0.1% in psoriasis treatment[2]
Once-daily mometasone furoate 0.1% cream outperformed twice-daily hydrocortisone valerate 0.2% cream in pediatric atopic dermatitis (p=0.008)[5]
Early topical hydrocortisone formulations (1955) laid foundation for later optimized steroids like hydrocortisone valerate[3]

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In in vivo animal models, Hydrocortisone 17-valerate demonstrates significant anti-inflammatory and antimitotic activity. A classic 1973 study using the tape-stripped hairless mouse skin model evaluated its efficacy, showing that a 1% concentration of the drug under occlusive dressing for 24 hours produced a three- to four-fold drop in the colcemid-induced mitotic index. The antimitotic effect started at or before 5 hours after administration and continued for at least 4 days after removal of the active material. The antimitotic effect of hydrocortisone 17-valerate was detectable at a concentration of 10⁻⁴, demonstrating significantly greater potency compared to hydrocortisone (which required 10⁻² concentration).

Glucocorticoid receptor binding assays typically employ a radioligand competitive binding method. The brief protocol is as follows: Prepare cytosol or recombinant receptor protein from GR-expressing cell lines or rat liver/thymus tissue. Mix a fixed concentration of radiolabeled ligand (e.g., ³H-dexamethasone, approximately 1-10 nM) with serial dilutions of Hydrocortisone 17-valerate (ranging from 10⁻¹⁰ to 10⁻⁵ M), then add the receptor protein and incubate at 4°C for 12-24 hours to reach equilibrium. Following incubation, adsorb unbound free ligand using dextran-coated charcoal (DCC) suspension, centrifuge, and measure radioactivity in the supernatant using a scintillation counter. Generate a competition binding curve by nonlinear regression analysis to calculate the IC₅₀ (concentration required to inhibit 50% of radioligand binding), and convert to the inhibition constant (Ki) using the Cheng-Prusoff equation.

Human T lymphocytes or peripheral blood mononuclear cells (PBMCs) are commonly used to evaluate the immunosuppressive activity of this drug. The standard protocol is as follows: Isolate PBMCs from healthy volunteer peripheral blood by Ficoll density gradient centrifugation, or use Jurkat T cell lines. Seed cells at a density of 1×10⁵ cells per well in a 96-well plate. Stimulate T cell activation by adding phytohemagglutinin (PHA, final concentration 5 μg/mL) or anti-CD3/CD28 antibodies, while adding serially diluted Hydrocortisone 17-valerate (typically dissolved in DMSO and diluted with culture medium to final concentrations of 0.01-100 μM). After incubation for 72 hours at 37°C in a 5% CO₂ incubator, add CCK-8 or MTT reagent to detect cell proliferation activity. Measure absorbance at 570 nm or 450 nm. Evaluate in vitro cellular activity by calculating the IC₅₀ for inhibition of lymphocyte proliferation.

Minimal systemic absorption: Plasma cortisol levels unchanged after 8g/day application for 7 days[1]
Trace amounts detected in urine (<1% of dose)[1]

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A classic in vivo pharmacodynamic evaluation uses the tape-stripped hairless mouse model. The specific protocol is as follows: Use hairless mice (e.g., hr/hr strain), repeatedly apply adhesive tape to the back skin to remove the stratum corneum, establishing a skin barrier-impaired inflammatory model. After anesthesia, soak small pieces of Whatman No. 1 filter paper in the drug solution (cream base containing 1% Hydrocortisone 17-valerate), apply to the stripped skin area, and secure with occlusive dressing. Remove the dressing after 24 hours of treatment, and collect samples at various time points (e.g., 5 hours, 24 hours, 2 days, 4 days). Administer colcemid (2 mg/kg) intraperitoneally 4 hours before sampling to arrest cells at metaphase. Prepare skin sections and count the number of mitotic cells per 1000 epidermal basal cells to calculate the mitotic index (MI). Evaluate efficacy by calculating the inhibition rate compared to the blank control group.

Absorption, Distribution and Excretion
Topical corticosteroids are absorbed through normal, intact skin. Skin inflammation and/or other conditions can increase percutaneous absorption. Corticosteroids are primarily metabolized in the liver and then excreted via the kidneys. Some topical corticosteroids and their metabolites are also excreted via bile. Metabolism/Metabolites Primarily metabolized in the liver via CYP3A4. Biological Half-Life 6-8 hours

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As a topical formulation, the percutaneous absorption of Hydrocortisone 17-valerate is determined by multiple factors, including vehicle properties, epidermal barrier integrity, application area, and the use of occlusive dressings. On intact skin, the drug exhibits limited percutaneous absorption; however, absorption significantly increases when the skin barrier is compromised (e.g., inflammation, skin diseases) or when occlusive dressings are used. One study indicates that occlusion for 24 hours does not increase penetration, but occlusion for 96 hours markedly enhances penetration. Once absorbed into circulation, the drug shares similar metabolic pathways with hydrocortisone, primarily undergoing hepatic metabolism, with metabolites excreted renally. Compared to topical formulations, PK parameters for oral or parenteral administration are less reported in the literature, as its clinical use is predominantly topical.

Protein Binding
95%
The toxicity of Hydrocortisone 17-valerate is primarily associated with systemic absorption. Long-term or widespread use may lead to reversible hypothalamic-pituitary-adrenal (HPA) axis suppression, manifesting as Cushing's syndrome, hyperglycemia, and glucosuria. The incidence of local adverse reactions is approximately 12%, including worsening of condition (2%), transient itching (2%), irritation (1%), and redness (1%). Pediatric patients have a higher incidence of adverse events (approximately 28.1%) due to their greater skin surface area-to-body mass ratio, making them more susceptible to HPA axis suppression and growth suppression. Long-term topical use may also cause skin atrophy, telangiectasia, hypopigmentation, acneiform eruptions, and folliculitis. Mutagenicity studies show that the drug is negative in the Ames test. Animal reproductive toxicity studies indicate embryotoxicity (e.g., decreased fetal weight, delayed skeletal ossification) and teratogenicity (e.g., cleft palate, omphalocele, clubbed feet) at high doses. No adequate and well-controlled studies exist in pregnant women.

[1]. Clinical evaluation of hydrocortisone valerate 0.2% ointment. Clin Ther. 1984;6(3):282-93.
[2]. Hydrocortisone valerate. Double-blind comparison with two other topical steroids. Cutis. 1978 May;21(5):695-8.
[3]. TOPICAL hydrocortisone. Br Med J. 1955 Feb 26;1(4912):530-1.
[4]. Comparative efficacy of hydrocortisone valerate 0.2 percent ointment in the treatment of atopic dermatitis. Cutis. 1983 Jul;32(1):89-91, 94.
[5]. A comparison of once-daily application of mometasone furoate 0.1% cream compared with twice-daily hydrocortisone valerate 0.2% cream in pediatric atopic dermatitis patients who failed to respond to hydrocortisone: mometasone furoate study group. Int J Dermatol. 1999 Aug;38(8):604-6.

17-Valtocortisol is a glucocorticoid, cortisol ester, valerate, and primary α-hydroxyketone. Hydrocortisone valerate is the valerate form of hydrocortisone and is a synthetic glucocorticoid receptor agonist with anti-inflammatory, antipruritic, and vasoconstrictive effects. Binding and activation of glucocorticoid receptors lead to the activation of lipocorticoids, which in turn inhibits cytosolic phospholipase A2. Deficiency of phospholipase A2 prevents the release of arachidonic acid (a precursor to the inflammatory mediators prostaglandins and leukotrienes) from the cell membrane. Secondly, mitogen-activated protein kinase (MAPK) phosphatase 1 is induced, leading to dephosphorylation and inactivation of the Jun N-terminal kinase, directly inhibiting c-Jun-mediated transcription. Finally, the transcriptional activity of nuclear factor (NF)-κB is blocked, thereby inhibiting the transcription of cyclooxygenase 2, which is essential for prostaglandin production. Pharmacological Indications: For the relief of inflammatory and pruritus symptoms in corticosteroid-responsive dermatitis. It is also used to treat endocrine (hormonal) disorders (adrenal insufficiency, Addison's disease). In addition, it is used to treat a variety of immune and allergic diseases, such as arthritis, lupus, severe psoriasis, severe asthma, ulcerative colitis, and Crohn's disease.

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Mechanism of Action
Hydrocortisone binds to cytoplasmic glucocorticoid receptors. After the newly formed receptor-ligand complex binds to the receptor, it translocates to the cell nucleus and binds to multiple glucocorticoid response elements (GREs) in the promoter regions of target genes. Subsequently, the DNA-bound receptor interacts with basal transcription factors, leading to increased expression of specific target genes. The anti-inflammatory effect of glucocorticoids is thought to be related to lipocortin, a protein that inhibits phospholipase A2, which controls the biosynthesis of prostaglandins and leukotrienes by inhibiting arachidonic acid. Specifically, glucocorticoids induce the synthesis of lipocortin-1 (annexin-1), which then binds to the cell membrane, preventing phospholipase A2 from contacting its substrate arachidonic acid. This leads to a decrease in the production of arachidic acid. The expression of cyclooxygenases (COX-1 and COX-2) is also inhibited, thereby enhancing the above effects. In other words, the two main products of inflammation—prostaglandins and leukotrienes—are inhibited by glucocorticoids. Glucocorticoids can also stimulate lipocortin-1 to escape into the extracellular space, where it binds to leukocyte membrane receptors, inhibiting various inflammatory responses: epithelial cell adhesion, migration, chemotaxis, phagocytosis, respiratory burst, and the release of various inflammatory mediators (lysosomal enzymes, cytokines, tissue plasminogen activator, chemokines, etc.) from neutrophils, macrophages, and mast cells. Furthermore, corticosteroids suppress the immune system through mechanisms including decreased lymphatic system function, reduced immunoglobulin and complement concentrations, lymphopenia, and interference with antigen-antibody binding.

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Pharmacodynamics
Hydrocortisone is the most important glucocorticoid in the human body. It is essential for life and regulates or supports many important cardiovascular, metabolic, immune, and homeostatic functions.
Topical application of hydrocortisone utilizes its anti-inflammatory or immunosuppressive properties to treat inflammation caused by corticosteroid-sensitive dermatitis. Glucocorticoids are a class of steroid hormones characterized by their ability to bind to cortisol receptors and trigger a variety of important cardiovascular, metabolic, immune, and homeostatic effects. Glucocorticoids differ from mineralocorticoids and sex hormones in that they have different receptors, target cells, and mechanisms of action. Strictly speaking, the term "corticosteroid" refers to both glucocorticoids and mineralocorticoids, but it is often used synonymously with glucocorticoids. Glucocorticoids suppress cell-mediated immune responses. They function by inhibiting genes encoding cytokines IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, and TNF-α, with IL-2 being the most important. Reduced cytokine production limits T cell proliferation. Glucocorticoids also suppress humoral immunity, leading to a decrease in IL-2 expression and its receptor on B cells. This reduces B cell clonal expansion and antibody synthesis. Decreased IL-2 levels also result in a reduction in the number of activated T lymphocytes.

C26H38O6

446.58

446.267

C, 69.93; H, 8.58; O, 21.50

57524-89-7

Hydrocortisone acetate;50-03-3;Hydrocortisone;50-23-7;Hydrocortisone phosphate;3863-59-0

5282494

White to off-white solid powder

1.21g/cm3

595.1ºC at 760mmHg

217-220 °C

195ºC

1.56

3.522

2

6

7

32

832

7

CCCCC(O[C@]1(C(CO)=O)CC[C@H]2[C@@H]3CCC4=CC(CC[C@@]4([C@H]3[C@@H](O)C[C@@]21C)C)=O)=O.C/C=C\C(C(NC1=CC=CC=C1C)=O)=C

FZCHYNWYXKICIO-FZNHGJLXSA-N

InChI=1S/C26H38O6/c1-4-5-6-22(31)32-26(21(30)15-27)12-10-19-18-8-7-16-13-17(28)9-11-24(16,2)23(18)20(29)14-25(19,26)3/h13,18-20,23,27,29H,4-12,14-15H2,1-3H3/t18-,19-,20-,23+,24-,25-,26-/m0/s1

[(8S,9S,10R,11S,13S,14S,17R)-11-hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-3-oxo-2,6,7,8,9,11,12,14,15,16-decahydro-1H-cyclopenta[a]phenanthren-17-yl] pentanoate

HYDROCORTISONE VALERATE; Cortisol 17-valerate; 57524-89-7; Hydrocortisone 17-valerate; Westcort; DTXSID9045633; CHEBI:50865; 68717P8FUZ;

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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.)