ABSTRACT
BACKGROUND: Dalcinonacog alfa (DalcA), a next-generation, recombinant human factor IX (FIX) variant, was developed using a rational design approach for increased procoagulant activity and longer duration of action to be administered subcutaneously (SC) for prophylaxis of hemophilia B bleeding episodes. OBJECTIVES: To investigate the safety, pharmacokinetics (PK), and pharmacodynamics (PD) of DalcA. METHODS: This multicenter, phase 1/2a study (NCT03186677) was conducted in 11 males aged 12 to 65 years with severe hemophilia B. In cohort 1, subjects received intravenous (IV) 75 IU/kg BeneFIX and DalcA. Cohorts 2 and 3 had DalcA IV 75 IU/kg and SC 75 IU/kg or 150 IU/kg. Cohort 4 was omitted. Cohort 5 received daily SC 150 IU/kg DalcA for 6 days and cohort 6 received IV 75 IU/kg and daily SC 150 IU/kg DalcA for 9 days. Blood sampling was performed for chemistry, hematology, PK, PD, and anti-drug antibody measurement. Subjects were monitored for safety endpoints for 30 days postdosing. RESULTS: DalcA demonstrated a 24-fold greater potency over BeneFIX and longer mean residence time (33.8 h). SC bioavailability 8.2% to 20.3%, beta half-life 53.9 to 106.9 h and Tmax 24 to 48 h. A median 15.7% FIX activity level (interquartile range, 14.9%-16.6%) was reached after 6 daily doses. Neutralizing antibodies to ISU304, but not wild-type FIX, occurred in two cousins. CONCLUSIONS: The data demonstrated that DalcA achieved protective FIX activity levels between 11% and 18%, corresponding to a reduced chance of spontaneous bleeds. Based on the results, a phase 2b trial to assess the safety and efficacy of 28 daily SC doses of DalcA was performed.
Subject(s)
Hemophilia A , Hemophilia B , Adolescent , Adult , Aged , Blood Coagulation Tests , Child , Factor IX , Half-Life , Hemophilia B/diagnosis , Hemophilia B/drug therapy , Humans , Male , Middle Aged , Recombinant Proteins , Young AdultABSTRACT
Apigenin (4',5,7-trihydroxyflavone, flavonoid) is a phenolic compound that is known to reduce the risk of chronic disease owing to its low toxicity. The first study on apigenin analyzed its effect on histamine release in the 1950s. Since then, anti-mutation and antitumor properties of apigenin have been widely reported. In the present study, we evaluated the apigenin-mediated amelioration of skin disease and investigated its applicability as a functional ingredient, especially in cosmetics. The effect of apigenin on RAW264.7 (murine macrophage), RBL-2H3 (rat basophilic leukemia), and HaCaT (human immortalized keratinocyte) cells were analyzed. Apigenin (100 µM) significantly inhibited nitric oxide (NO) production, cytokine expression (interleukin (IL)-1ß, IL6, cyclooxygenase (COX)-2, and inducible nitric oxide synthase [iNOS]), and phosphorylation of mitogen-activated protein kinase (MAPK) signal molecules, including extracellular signal-regulated kinase (ERK) and c-Jun N-terminal protein kinase (JNK) in RAW264.7 cells. Apigenin (30 M) also inhibited the phosphorylation of signaling molecules (Lyn, Syk, phospholipase Cγ1, ERK, and JNK) and the expression of high-affinity IgE receptor FcεRIα and cytokines (tumor necrosis factor (TNF)-α, IL-4, IL-5, IL-6, IL-13, and COX-2) that are known to induce inflammation and allergic responses in RBL-2H3 cells. Further, apigenin (20 µM) significantly induced the expression of filaggrin, loricrin, aquaporin-3, hyaluronic acid, hyaluronic acid synthase (HAS)-1, HAS-2, and HAS-3 in HaCaT cells that are the main components of the physical barrier of the skin. Moreover, it promoted the expression of human ß-defensin (HBD)-1, HBD-2, HBD-3, and cathelicidin (LL-37) in HaCaT cells. These antimicrobial peptides are known to play an important role in the skin as chemical barriers. Apigenin significantly suppressed the inflammatory and allergic responses of RAW264.7 and RBL cells, respectively, and would, therefore, serve as a potential prophylactic and therapeutic agent for immune-related diseases. Apigenin could also be used to improve the functions of the physical and chemical skin barriers and to alleviate psoriasis, acne, and atopic dermatitis.
Subject(s)
Apigenin/pharmacology , Skin Diseases/drug therapy , Animals , Anti-Allergic Agents/metabolism , Anti-Allergic Agents/pharmacology , Anti-Inflammatory Agents/metabolism , Anti-Inflammatory Agents/pharmacology , Apigenin/metabolism , Cell Line , Cytokines/metabolism , Extracellular Signal-Regulated MAP Kinases/metabolism , Filaggrin Proteins , HaCaT Cells/drug effects , Humans , Immunoglobulin E/metabolism , Interleukin-1beta/metabolism , JNK Mitogen-Activated Protein Kinases/metabolism , Mast Cells/metabolism , Mice , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II/metabolism , Phosphorylation/drug effects , RAW 264.7 Cells/drug effects , Rats , Receptors, IgE/genetics , Signal Transduction/drug effects , Skin/drug effects , Skin/metabolism , Skin Physiological Phenomena/drug effects , Tumor Necrosis Factor-alpha/metabolismABSTRACT
In this study, recombinant human bone morphogenetic protein-2 (rhBMP-2) was directly immobilized on the plasma-polymerized propionaldehyde (PA) and allylglycidyl ether (AGE) surface through the imine bonding and epoxy-amine bonding, respectively. Aldehyde and epoxide plasma-polymerization were carried out at plasma power 60 W for 10 min and monomers were used to PA and AGE. After the plasma-polymerization and rhBMP-2 immobilization, substrate surfaces were characterized by contact angle, field emission scanning electron microscopy, and attenuated total reflectance Fourier transform infrared. In addition, the biological activities of MC3T3-E1 cells were evaluated by initial adhesion and alkaline phosphate (ALP) activity. The rhBMP-2 immobilized PA and AGE surfaces promoted significantly higher ALP activity of MC3T3-E1 cells than pristine surface.
