K252b

Last updated
K252b
K252b.svg
Names
IUPAC name
(15S,16R,18R)-16-Hydroxy-15-methyl-3-oxo-28-oxa-4,14,19-triazaoctacyclo[12.11.2.115,18.02,6.07,27.08,13.019,26.020,25]octacosa-1,6,8,10,12,20,22,24,26-nonaene-16-carboxylic acid
Identifiers
3D model (JSmol)
ChEBI
ChEMBL
ChemSpider
PubChem CID
  • InChI=1S/C26H19N3O5/c1-25-26(33,24(31)32)10-17(34-25)28-15-8-4-2-6-12(15)19-20-14(11-27-23(20)30)18-13-7-3-5-9-16(13)29(25)22(18)21(19)28/h2-9,17,33H,10-11H2,1H3,(H,27,30)(H,31,32)/t17-,25+,26+/m1/s1
    Key: AMSOPBXQXSAAAC-PLZPTFKGSA-N
  • C[C@@]12[C@](C[C@@H](O1)N3C4=CC=CC=C4C5=C6C(=C7C8=CC=CC=C8N2C7=C53)CNC6=O)(C(=O)O)O
Properties [1]
C26H19N3O5
Molar mass 453.454 g·mol−1
AppearanceWhite to light yellow powder
Solubility soluble in DMSO, methanol
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).

K252b is an ectoprotein kinase inhibitor, which is involved in the abolishment of the effects of nerve growth factors on PC12 and peripheral neuron system (PNS) neurons. When it is present in very low concentrations, it prolongs the survivorship of hippocampal, septal and cortical neurons deprived of glucose. [2]

Contents

K252b is related to K252a and staurosporine, which are low-molecular-weight alkaloids. [3] Staurosporine was discovered in 1977 while screening for microbial alkaloids using chemical detection methods. K252 was discovered in 1986 as a related natural indolocarbazole product. In 2007, K252b was found to have an inhibitory effect on mycobacterial protein kinase PknB. This is a receptor-like transmembrane protein. The PknB gene can be found in all known mycobacterial genomes. Staurosporine and K252a have inhibitory effects on the growth of Mycobacterium tuberculosis , but K252b failed to inhibit bacterial growth.

Metabolism

The metabolism of K252b, a complex organic molecule, is caused by interactions between the molecule and various metabolic pathways in different living organisms, [4] microorganisms and animals are among these organisms.

Uses

In influencing NGF and neurite outgrowth

K252b controls neurite formation by inhibiting Nerve Growth Factor(NGF)-induced neurite outgrowth of PC12 cells. [5] [6] This inhibition is dose-dependent with higher concentrations of K252b leading to more NGF-induced neurite outgrowth inhibition. The exact mechanism of K252b inhibition of NGF-induced neurite outgrowth is still largely unknown. [5] [6]

In synapse formation

Continuous application of K252b also dose-dependently inhibits the formation of synapses in the cultured cortical neurons of rat brains, with higher doses resulting in higher inhibition. K252b does this by suppressing phosphorylation of the extracellular domains of proteins, mainly the microtubule-associated protein (MAP) 1B is sensitive to the inhibiting effects of K252b. [7] Additionally, K252b application causes a significant drop in the frequency of synchronous firing of cultured cortical neurons of rats. [7]

In immune response processes

K252b inhibits the immune response of mast cells and human basophil cells in several ways. Inhibition of the mast cell and human basophil immune response by involvement of ectokinase K252b in IgE-receptor mediated signaling has been speculated to function as a suppressor of acute allergic reactions. [8] K252b's inhibition is dose-dependent, with higher concentrations of K252b resulting in more inhibition. The role of K252b in suppressing the immune response lies mainly in preventing cytosolic Ca2+ increase and preventing degranulation and histamine release in mast cells and human basophils. [8]

Effects of K252b on cytosolic Ca2+ concentrations

K252b inhibits Ca2+ cytosolic concentration increases. K252b does this by inhibiting the Ca2+ influx from extracellular fluid and Ca2+ release from intracellular storage, therefore preventing the increase of the Ca2+ cytosolic concentration, which is an essential step in the process of activation of the immune response of mast cells and human basophil cells. [8]

Effects on degranulation and histamine release of mast cells and human basophils

K252b inhibits basophil and mast-cell histamine release induced by both Ag and BA3. Additionally, K252b dose-dependently inhibits Ag induced β-hexosaminidase release. [8] K252b also inhibits IgE receptor-mediated degranulation of mast cells, in this process K252b acts at a very early stage of IgE receptor-mediated stimulation, which according to Teshima et al. means that the target for K252b is likely an early signaling molecule active in the process of IgE receptor mediation. [8] K252b also suppresses the IgE receptor-mediated histamine release and to a lower extent also 150 nM TPA mediated histamine release in mast cells and human basophil cells. Degranulation is, like increase in the cytosolic Ca2+ concentration, an essential step in the activation of the immune response of mast cells and human basophil cells. [8]

Efficacy

In general

K252b has been shown to have a higher efficacy at higher doses, making its inhibiting processes dose dependent. [5] [7] [8] Since K252b does not permeate the cell membrane of the cells it acts upon, due to its hydrophilic nature, K252b generally has a low cytotoxicity and leaves less cytotoxic damage than its membrane-permeating analogs like K252a. [5] [7] [8]

In the process of NGF-induced neurite outgrowth inhibition

K252b has been shown to entirely inhibit NGF-induced neurite outgrowth at concentrations of 300 nM, while K252b concentrations of 100 nM or less result in a partial inhibition. [5]

Toxicology data

The CC50 was reported to be less than 40 μM against murine macrophage J774A.1 measured by AlamarBlue assay. [9]

Structure, reactivity, and synthesis

K252b is a large molecule which falls into the indolocarbozole category. K252b looks very much like another indolocarbazole: K252a. The difference between these 2 is that K252b possesses a carboxylic acid group where K252a has an ester instead. This difference makes K252b a lot more hydrophobic than K252a, the partition coefficient for K252b is 4.4:1 (org:aq) and 15.6:1 for K252a2. [4] This difference in hydrophilicity has an impact on the uptake and concentration of K252b into cells. K252b is taken up reversibly by PC12 and Sf9 cells, whereas K252a is taken up irreversibly at a higher concentration. [4]

One path to synthesize K252b, would be to synthesize K252a first and then react K252a with a strong base. K252a has been synthesized in many papers, [10] there is no known route to directly synthesize K252b yet.

Molecular mechanism of action

K252b inhibits tyrosine kinase receptors and the long-term trophic effects of NT-3 and BDNF at concentrations above 100 nM. [11] At concentrations under 100 nM it stimulates the trophic effects of NT-3. [11]

See also

Related Research Articles

<span class="mw-page-title-main">Mast cell</span> Cell found in connective tissue

A mast cell is a resident cell of connective tissue that contains many granules rich in histamine and heparin. Specifically, it is a type of granulocyte derived from the myeloid stem cell that is a part of the immune and neuroimmune systems. Mast cells were discovered by Paul Ehrlich in 1877. Although best known for their role in allergy and anaphylaxis, mast cells play an important protective role as well, being intimately involved in wound healing, angiogenesis, immune tolerance, defense against pathogens, and vascular permeability in brain tumors.

<span class="mw-page-title-main">Basophil</span> Type of white blood cell

Basophils are a type of white blood cell. Basophils are the least common type of granulocyte, representing about 0.5% to 1% of circulating white blood cells. However, they are the largest type of granulocyte and how they work is not fully understood. They are responsible for inflammatory reactions during immune response, as well as in the formation of acute and chronic allergic diseases, including anaphylaxis, asthma, atopic dermatitis and hay fever. They also produce compounds that coordinate immune responses, including histamine and serotonin that induce inflammation, and heparin that prevents blood clotting, although there are less than that found in mast cell granules. Mast cells were once thought to be basophils that migrated from the blood into their resident tissues, but they are now known to be different types of cells.

<span class="mw-page-title-main">Immunoglobulin E</span> Immunoglobulin E (IgE) Antibody

Immunoglobulin E (IgE) is a type of antibody that has been found only in mammals. IgE is synthesised by plasma cells. Monomers of IgE consist of two heavy chains and two light chains, with the ε chain containing four Ig-like constant domains (Cε1–Cε4). IgE is thought to be an important part of the immune response against infection by certain parasitic worms, including Schistosoma mansoni, Trichinella spiralis, and Fasciola hepatica. IgE is also utilized during immune defense against certain protozoan parasites such as Plasmodium falciparum. IgE may have evolved as a defense to protect against venoms.

<span class="mw-page-title-main">Type I hypersensitivity</span> Type of allergic reaction

Type I hypersensitivity, in the Gell and Coombs classification of allergic reactions, is an allergic reaction provoked by re-exposure to a specific type of antigen referred to as an allergen. Type I is distinct from type II, type III and type IV hypersensitivities. The relevance of the Gell and Coombs classification of allergic reactions has been questioned in the modern-day understanding of allergy, and it has limited utility in clinical practice.

Ryanodine receptors form a class of intracellular calcium channels in various forms of excitable animal tissue like muscles and neurons. There are three major isoforms of the ryanodine receptor, which are found in different tissues and participate in different signaling pathways involving calcium release from intracellular organelles. The RYR2 ryanodine receptor isoform is the major cellular mediator of calcium-induced calcium release (CICR) in animal cells.

<span class="mw-page-title-main">Nerve growth factor</span> Mammalian protein found in Homo sapiens

Nerve growth factor (NGF) is a neurotrophic factor and neuropeptide primarily involved in the regulation of growth, maintenance, proliferation, and survival of certain target neurons. It is perhaps the prototypical growth factor, in that it was one of the first to be described. Since it was first isolated by Nobel Laureates Rita Levi-Montalcini and Stanley Cohen in 1956, numerous biological processes involving NGF have been identified, two of them being the survival of pancreatic beta cells and the regulation of the immune system.

Histamine H<sub>2</sub> receptor Mammalian protein found in Homo sapiens

H2 receptors are positively coupled to adenylate cyclase via Gs alpha subunit. It is a potent stimulant of cAMP production, which leads to activation of protein kinase A. PKA functions to phosphorylate certain proteins, affecting their activity. The drug betazole is an example of a histamine H2 receptor agonist.

<span class="mw-page-title-main">K252a</span> Chemical compound

K252a is an alkaloid isolated from Nocardiopsis bacteria. This staurosporine analog is a highly potent cell permeable inhibitor of CaM kinase and phosphorylase kinase (IC50 = 1.8 and 1.7 nmol/L, respectively). At higher concentrations it is also an efficient inhibitor of serine/threonine protein kinases (IC50 of 10 to 30 nmol/L).

A neurite or neuronal process refers to any projection from the cell body of a neuron. This projection can be either an axon or a dendrite. The term is frequently used when speaking of immature or developing neurons, especially of cells in culture, because it can be difficult to tell axons from dendrites before differentiation is complete.

<span class="mw-page-title-main">FCER1</span>

The high-affinity IgE receptor, also known as FcεRI, or Fc epsilon RI, is the high-affinity receptor for the Fc region of immunoglobulin E (IgE), an antibody isotype involved in allergy disorders and parasite immunity. FcεRI is a tetrameric receptor complex that binds Fc portion of the ε heavy chain of IgE. It consists of one alpha, one beta, and two gamma chains connected by two disulfide bridges on mast cells and basophils. It lacks the beta subunit on other cells. It is constitutively expressed on mast cells and basophils and is inducible in eosinophils.

<span class="mw-page-title-main">Tropomyosin receptor kinase A</span> Protein-coding gene in the species Homo sapiens

Tropomyosin receptor kinase A (TrkA), also known as high affinity nerve growth factor receptor, neurotrophic tyrosine kinase receptor type 1, or TRK1-transforming tyrosine kinase protein is a protein that in humans is encoded by the NTRK1 gene.

<span class="mw-page-title-main">Lipid signaling</span> Biological signaling using lipid molecules

Lipid signaling, broadly defined, refers to any biological cell signaling event involving a lipid messenger that binds a protein target, such as a receptor, kinase or phosphatase, which in turn mediate the effects of these lipids on specific cellular responses. Lipid signaling is thought to be qualitatively different from other classical signaling paradigms because lipids can freely diffuse through membranes. One consequence of this is that lipid messengers cannot be stored in vesicles prior to release and so are often biosynthesized "on demand" at their intended site of action. As such, many lipid signaling molecules cannot circulate freely in solution but, rather, exist bound to special carrier proteins in serum.

<span class="mw-page-title-main">Myelin-associated glycoprotein</span> Protein-coding gene in the species Homo sapiens

Myelin-associated glycoprotein is a type 1 transmembrane protein glycoprotein localized in periaxonal Schwann cell and oligodendrocyte membranes, where it plays a role in glial-axonal interactions. MAG is a member of the SIGLEC family of proteins and is a functional ligand of the NOGO-66 receptor, NgR. MAG is believed to be involved in myelination during nerve regeneration in the PNS and is vital for the long-term survival of the myelinated axons following myelinogenesis. In the CNS MAG is one of three main myelin-associated inhibitors of axonal regeneration after injury, making it an important protein for future research on neurogenesis in the CNS.

<span class="mw-page-title-main">Staurosporine</span> Chemical compound

Staurosporine is a natural product originally isolated in 1977 from the bacterium Streptomyces staurosporeus. It was the first of over 50 alkaloids to be isolated with this type of bis-indole chemical structure. The chemical structure of staurosporine was elucidated by X-ray analysis of a single crystal and the absolute stereochemical configuration by the same method in 1994.

<span class="mw-page-title-main">Degranulation</span> Process by which cells lose secretory granules

Degranulation is a cellular process that releases antimicrobial cytotoxic or other molecules from secretory vesicles called granules found inside some cells. It is used by several different cells involved in the immune system, including granulocytes. It is also used by certain lymphocytes such as natural killer (NK) cells and cytotoxic T cells, whose main purpose is to destroy invading microorganisms.

Trk receptors are a family of tyrosine kinases that regulates synaptic strength and plasticity in the mammalian nervous system. Trk receptors affect neuronal survival and differentiation through several signaling cascades. However, the activation of these receptors also has significant effects on functional properties of neurons.

<span class="mw-page-title-main">SIGLEC8</span> Protein-coding gene in the species Homo sapiens

Sialic acid-binding Ig-like lectin 8 is a protein that in humans is encoded by the SIGLEC8 gene. This gene is located on chromosome 19q13.4, about 330 kb downstream of the SIGLEC9 gene. Within the siglec family of transmembrane proteins, Siglec-8 belongs to the CD33-related siglec subfamily, a subfamily that has undergone rapid evolution.

<span class="mw-page-title-main">Basophil activation</span>

Allergic symptoms are caused by an initial systemic histamine release by activated basophils and mast cells, that may lead to shock with laryngeal edema, lower-airway obstruction and hypotension. This is why basophils are considered with mast cells to be the key cells in allergic diseases.

N2a cells are a fast-growing mouse neuroblastoma cell line.

Phosphatidylinositol-4-phosphate 5-kinases are a class of enzymes that phosphorylate phosphatidylinositol 4-phosphate. They perform this reaction on the fifth hydroxyl of the myo-inositol ring to form phosphatidylinositol 4,5-bisphosphate.

References

  1. "K252b | Fermentek". www.fermentek.com. Retrieved 13 March 2024.
  2. Cheng, Bin; Barger, Steven W.; Mattson, Mark P. (April 1994). "Staurosporine, K-252a, and K-252b Stabilize Calcium Homeostasis and Promote Survival of CNS Neurons in the Absence of Glucose". Journal of Neurochemistry. 62 (4): 1319–1329. doi:10.1046/j.1471-4159.1994.62041319.x. ISSN   0022-3042. PMID   7510777. S2CID   19294051.
  3. Nakano, Hirofumi; Ōmura, Satoshi (January 2009). "Chemical biology of natural indolocarbazole products: 30 years since the discovery of staurosporine". The Journal of Antibiotics. 62 (1): 17–26. doi:10.1038/ja.2008.4. ISSN   1881-1469. PMID   19132059.
  4. 1 2 3 Ross, Alonzo H.; McKinnon, Christine A.; Daou, Marie-Claire; Ratliff, Kristin; Wolf, David E. (December 1995). "Differential Biological Effects of K252 Kinase Inhibitors Are Related to Membrane Solubility but Not to Permeability". Journal of Neurochemistry. 65 (6): 2748–2756. doi:10.1046/j.1471-4159.1995.65062748.x. ISSN   0022-3042. PMID   7595574. S2CID   26026172.
  5. 1 2 3 4 5 Nagashima, Ken; Nakanishi, Satoshi; Matsuda, Yuzuru (November 1991). "Inhibition of nerve growth factor-induced neurite outgrowth of PC12 cells by a protein kinase inhibitor which does not permeate the cell membrane". FEBS Letters. 293 (1–2): 119–123. doi:10.1016/0014-5793(91)81165-5. ISSN   0014-5793. PMID   1959642.
  6. 1 2 Knüsel, Beat; Hefti, Franz (September 1991). "K-252b Is a Selective and Nontoxic Inhibitor of Nerve Growth Factor Action on Cultured Brain Neurons". Journal of Neurochemistry. 57 (3): 955–962. doi:10.1111/j.1471-4159.1991.tb08243.x. ISSN   0022-3042. PMID   1861160. S2CID   36087741.
  7. 1 2 3 4 Muramoto, K.; Taniguchi, H.; Kawahara, M.; Kobayashi, K.; Nonomura, Y.; Kuroda, Y. (1994-12-15). "A Substrate of Ecto-Protein Kinase Is Microtubule-Associated Protein 1B in Cortical Cell Cultures Undergoing Synaptogenesis". Biochemical and Biophysical Research Communications. 205 (2): 1467–1473. doi:10.1006/bbrc.1994.2830. ISSN   0006-291X. PMID   7802683.
  8. 1 2 3 4 5 6 7 8 Teshima, R; Saito, Y; Ikebuchi, H; Rajiva De Silva, N; Morita, Y; Nakanishi, M; Sawada, J; Kitani, S (1997-07-15). "Effect of an ectokinase inhibitor, K252b, on degranulation and Ca2+ signals of RBL-2H3 cells and human basophils". The Journal of Immunology. 159 (2): 964–969. doi:10.4049/jimmunol.159.2.964. ISSN   0022-1767. PMID   9218617. S2CID   11866386.
  9. Cartuche, Luis; Sifaoui, Ines; López-Arencibia, Atteneri; Bethencourt-Estrella, Carlos J.; San Nicolás-Hernández, Desirée; Lorenzo-Morales, Jacob; Piñero, José E.; Díaz-Marrero, Ana R.; Fernández, José J. (April 2020). "Antikinetoplastid Activity of Indolocarbazoles from Streptomyces sanyensis". Biomolecules. 10 (4): 657. doi: 10.3390/biom10040657 . ISSN   2218-273X. PMC   7226613 . PMID   32344693.
  10. Wood, John L.; Stoltz, Brian M.; Dietrich, Hans-Juergen (October 1995). "Total Synthesis of (+)- and (-)-K252a". Journal of the American Chemical Society. 117 (41): 10413–10414. doi:10.1021/ja00146a039. ISSN   0002-7863.
  11. 1 2 Plomp, J J; Molenaar, P C (1996-05-15). "Involvement of protein kinases in the upregulation of acetylcholine release at endplates of alpha-bungarotoxin-treated rats". The Journal of Physiology. 493 (1): 175–186. doi:10.1113/jphysiol.1996.sp021373. ISSN   0022-3751. PMC   1158959 . PMID   8735703.