PAR-1

On March 18th, Cell published online a research paper about a new mechanism involving the gut in regulating cholesterol metabolism. The researchers discovered a gut-derived hormone called Cholesin, which is triggered by cholesterol intake and produced within intestinal cells. Cholesin achieves inhibition of the PKA-ERK1/2 signaling pathway in the liver by binding to the GPR146 receptor there, thereby downregulating the cholesterol synthesis process regulated by SREBP2. This means that when the gut absorbs cholesterol, the Cholesin-GPR146 axis acts to suppress cholesterol production in the liver, helping to maintain stable blood cholesterol levels and combat hypercholesterolemia and atherosclerosis.  DOI: 10.1016/j.cell.2024.02.024To investigate the interaction between intestinal cholesterol absorption and hepatic cholesterol synthesis, the researchers designed experiments to induce mice to consume diets with varying cholesterol content (regular diet and high-cholesterol Western diet) and observed changes in cholesterol levels in the intestine and liver. Under a high-cholesterol diet, intestinal cholesterol content increased, accompanied by downregulated expression of the cholesterol biosynthesis marker HMGCR in the intestine, suggesting a negative feedback mechanism where absorbed cholesterol can influence its own synthesis. Against this backdrop, the researchers identified a new gut-derived hormone induced by cholesterol – Cholesin. Cholesin is triggered by cholesterol absorption mediated by NPC1L1, with its gene encoded in humans as C7orf50 and in mice as 3110082I17Rik. After secretion by intestinal cells, Cholesin binds to a specific orphan GPCR member, GPR146, and suppresses the PKA-ERK1/2 signaling pathway, reducing the hepatic cholesterol synthesis activity regulated by SREBP2. To further confirm the relationship between Cholesin and its receptor GPR146, researchers employed a variety of experimental approaches, including but not limited to binding assays on frozen tissue sections, genome-wide CRISPR-Cas9 screenings, and immunohistochemistry techniques. Ultimately, it was established that GPR146 is an effective receptor for Cholesin, and the interaction between Cholesin and GPR146 directly influences the regulation of cholesterol metabolism. Through CRISPR screenings, protein expression purification, and a range of experimental validations, Cholesin's mechanism of action was elucidated, and it was demonstrated that both the sole use of Cholesin and its combination with statin drugs significantly reduce total plasma cholesterol levels and alleviate atherosclerotic lesions. Moreover, Cholesin can also decrease body weight gain, lower triglyceride levels, and improve hepatic lipid accumulation and inflammation markers. About AtherosclerosisAtherosclerosis is a chronic inflammatory disease, primarily characterized by the accumulation of substantial amounts of cholesterol and other lipid substances on the vessel walls, forming plaques. These plaques gradually enlarge and can lead to the narrowing of the arterial lumen, impeding blood flow. Additionally, these plaques may become unstable and rupture, causing thrombosis, which in turn can trigger serious events such as myocardial infarction or stroke. Cholesterol is an essential lipid for the human body, involved in the construction of cell membranes, hormone synthesis, and various other physiological functions. However, when cholesterol levels in the blood are too high, especially when low-density lipoprotein cholesterol (LDL-C) concentrations are excessive, the risk for cardiovascular diseases increases, with the most notable being the onset and progression of atherosclerosis. Studies have shown that an excess of cholesterol, particularly oxidized LDL cholesterol, penetrating beneath endothelial cells, can trigger an inflammatory response through a complex series of biochemical processes. This stimulates smooth muscle cell migration and proliferation, causes macrophages to engulf lipids and become foam cells, forming what are called fatty streaks, which then evolve into mature atherosclerotic plaques. In addition to the abnormal accumulation of lipids, the state of imbalance between immune responses and their clearance is also believed to be primarily shaped by the migration and homeostasis of leukocytes, which are regulated by chemokines and their receptors. In recent years, new pro-inflammatory and anti-inflammatory pathways that link lipid biology with inflammation have been discovered, and gene expression profiling studies have revealed various factors involved in human coronary artery disease. Cardiovascular diseases are the leading cause of death globally, with an estimated 17.8 million people dying each year. Inflammation is considered to be one of the main driving forces in the development of atherosclerosis and is recognized as one of the major etiologies of cardiovascular disease. Despite the increasing body of research evidence over the past three decades showing that immune components play an indispensable role in the formation and chronicity of atherosclerosis, through a series of immunotherapeutic targets, the benefits and challenges brought about by targeting inflammation and the immune system in cardiovascular diseases have been demonstrated. DOI: 10.1038/s41569-021-00668-4DOI: 10.1038/nm.2538Using the diagram above as an example, chemokines and their receptors also play an important role in the pathogenesis of atherosclerosis. The process of leukocyte recruitment includes rolling, firm adhesion, lateral migration, and transendothelial migration, which are finely regulated by chemokines. On the one hand, soluble chemokines directly mediate the recruitment of leukocytes; on the other hand, chemokines immobilized by glycosaminoglycans (GAGs) on the surface of activated endothelial cells can trigger the docking of leukocytes through GPCRs and activate leukocyte integrins, thus achieving firm adhesion. The cell-specific and lesion stage-specific utilization of chemokines and their receptors underscores the robustness and complexity of the chemokine system, ensuring the formation of specific chemokine combinations at each stage of the lesion that attract specific subtypes of leukocytes. Synergistic effects among chemokines in the recruitment of leukocytes can lead to cooperative interactions, potentially mediating the process between leukocytes and atherosclerotic lesions. According to statistics from the Synapse database, as of now, there are more than 500 therapeutic drug pipelines globally, originating from over 400 institutions, encompassing over 200 targets, and involving 4095 clinical trials. The therapeutic drugs for this disease have a diverse range of Mechanisms of Action (MOA). At present, the primary therapeutic drugs for the treatment of atherosclerosis include statins, which function primarily by inhibiting the key enzyme in cholesterol biosynthesis—hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase. Examples include Lovastatin (Merck, approved in 1987), Fluvastatin (Novartis, approved in 1993), Atorvastatin (Pfizer, approved in 1996), Rosuvastatin (AstraZeneca, approved in 2002). P2Y12 receptor antagonists are another class of therapeutic drugs for atherosclerosis, acting by selectively blocking the function of the P2Y12 receptor to inhibit excessive activation and aggregation of platelets. This is crucial because platelets play a key role in the development of atherosclerosis and thrombosis, especially in the progression of cardiovascular diseases. Common drugs in this category include Clopidogrel (Sanofi, approved in 1997), Ticagrelor (AstraZeneca, approved in 2010), Cangrelor (CHIESI, approved in 2015). Additionally, in recent years, PCSK9 has emerged as a therapeutic candidate for the treatment of hypercholesterolemia and atherosclerosis-related diseases. The primary function of PCSK9 is to regulate the number of low-density lipoprotein cholesterol (LDL-C) receptors on the membrane of liver cells. When PCSK9 binds to LDL receptors, it leads to their degradation within lysosomes, thereby reducing the uptake and clearance of circulating LDL-C by the liver, resulting in elevated levels of LDL-C in the blood. Therapeutic drugs related to this mechanism include Evolocumab (Amgen, approved in 2015), Alirocumab (Sanofi, approved in 2015), Inclisiran (Novartis, approved in 2020). Factor Xa inhibitors are a class of drugs that specifically inhibit the activity of coagulation factor Xa (FXa). FXa plays a crucial role in the blood coagulation process, as it acts as a key enzyme in the coagulation cascade, facilitating the conversion of prothrombin to thrombin, which in turn catalyzes the transformation of fibrinogen into fibrin, leading to the formation of a stable clot. By binding to FXa and preventing its activity, Factor Xa inhibitors can effectively suppress the formation of thrombi and are widely used in clinical practice to prevent and treat various thrombotic diseases, such as deep vein thrombosis, pulmonary embolism, and stroke prevention in patients with atrial fibrillation. Therapeutic candidates in this category include Rivaroxaban (Bayer, approved in 2008), Edoxaban Tosylate (Daiichi Sankyo, approved in 2012), Apixaban (Bristol-Myers Squibb, approved in 2011). Additionally, SGLT2 inhibitors were initially developed for the treatment of type 2 diabetes by inhibiting the SGLT2 transporter protein in the renal proximal convoluted tubules, reducing the reabsorption of glucose and thereby promoting the excretion of glucose in the urine, leading to decreased blood glucose levels. In recent years, multiple large-scale clinical trials have revealed the significant therapeutic effect of SGLT2 inhibitors in reducing the risk of cardiovascular events, especially in patients with type 2 diabetes who have atherosclerotic cardiovascular diseases. Therapeutic candidates related to this include Tofogliflozin (Kowa, approved in 2014) and Ertugliflozin (Pfizer, approved in 2017). Besides the classical P2Y12 receptor, potential therapeutic targets within the GPCR family include AT1R, PAR-1R, CXCR4, CCR2, NIACR1, TBXA2R, PGI2R, ADRA1, and EP3. However, the progress in the drug pipeline related to these targets is not promising. Only Vorapaxar, a tricyclic compound derived from camptothecin that acts as a selective inhibitor on thrombin-activated receptor (PAR-1), has been approved for reducing the incidence of thrombotic cardiovascular events in patients with a history of myocardial infarction (MI) or peripheral artery disease (PAD). The first antibody drug candidate developed targeting an immunomodulatory factor associated with GPCRs for treating atherosclerosis is the humanized monoclonal antibody Plozalizumab, an antagonistic monoclonal antibody targeting CCR2 developed by Takeda Pharmaceutical in Japan. CCR2 plays a critical role in the migration of monocytes and macrophages to sites of inflammation. The recruitment of macrophages to the arterial wall is considered a key step in the development of atherosclerosis. The first nanobody drug candidate is BI-655088, targeting the CX3CR1 receptor.

Scientists show for the first time that bacteria can cause itch by activating nerve cells in the skin. The findings can inform new therapies to treat itch that occurs in inflammatory skin conditions like eczema and dermatitis. Scientists at Harvard Medical School have shown for the first time that a common skin bacterium -- Staphylococcus aureus -- can cause itch by acting directly on nerve cells. The findings, based on research in mice and in human cells, are reported Nov. 22 in Cell. The research adds an important piece to the long-standing puzzle of itch and helps explain why common skin conditions like eczema and atopic dermatitis are often accompanied by persistent itch. In such conditions, the equilibrium of microorganisms that keep our skin healthy is often thrown off balance, allowing S. aureus to flourish, the researchers said. Up until now, the itch that occurs with eczema and atopic dermatitis was believed to arise from the accompanying inflammation of the skin. But the new findings show that S. aureus single-handedly causes itch by instigating a molecular chain reaction that culminates in the urge to scratch. "We've identified an entirely novel mechanism behind itch -- the bacterium Staph aureus, which is found on almost every patient with the chronic condition atopic dermatitis. We show that itch can be caused by the microbe itself," said senior author Isaac Chiu, associate professor of immunology in the Blavatnik Institute at HMS. The study experiments showed that S. aureus releases a chemical that activates a protein on the nerve fibers that transmit signals from the skin to the brain. Treating animals with an FDA-approved anti-clotting medicine successfully blocked the activation of the protein to interrupt this key step in the itch-scratch cycle. The treatment relieved symptoms and minimized skin damage. The findings can inform the design of oral medicines and topical creams to treat persistent itch that occurs with various conditions linked to an imbalance in the skin microbiome, such as atopic dermatitis, prurigo nodularis, and psoriasis. The repeated scratching that is a hallmark of these conditions can cause skin damage and amplify inflammation. "Itch can be quite debilitating in patients who suffer from chronic skin conditions. Many of these patients carry on their skin the very microbe we've now shown for the first time can induce itch," said study first author Liwen Deng, a postdoctoral research fellow in the Chiu Lab. Identifying the molecular spark plug that ignites itch Researchers exposed the skin of mice to S. aureus. The animals developed intensifying itch over several days, and the repeated scratching caused worsening skin damage that spread beyond the original site of exposure. Moreover, mice exposed to S. aureus became hypersensitive to innocuous stimuli that would not typically cause itch. The exposed mice were more likely than unexposed mice to develop abnormal itching in response to a light touch. This hyperactive response, a condition called alloknesis, is common in patients with chronic conditions of the skin characterized by persistent itch. But it can also happen in people without any underlying conditions -- think of that scratchy feeling you might get from a wool sweater. To determine how the bacterium triggered itch, the researchers tested multiple modified versions of the S. aureus microbe that were engineered to lack specific pieces of the bug's molecular makeup. The team focused on 10 enzymes known to be released by this microbe upon skin contact. One after another, the researchers eliminated nine suspects -- showing that a bacterial enzyme called protease V8 was single-handedly responsible for initiating itch in mice. Human skin samples from patients with atopic dermatitis also had more S. aureus and higher V8 levels than healthy skin samples. The analyses showed that V8 triggers itch by activating a protein called PAR1, which is found on skin neurons that originate in the spinal cord and carry various signals -- touch, heat, pain, itch -- from the skin to the brain. Normally, PAR1 lies dormant but upon contact with certain enzymes, including V8, it gets activated. The research showed that V8 snips one end of the PAR1 protein and awakens it. Experiments in mice showed that once activated, PAR1 initiates a signal that the brain eventually perceives as itch. When researchers repeated the experiments in lab dishes containing human neurons, they also responded to V8. Interestingly, various immune cells implicated in skin allergies and classically known to cause itch -- mast cells and basophils -- did not drive itch after bacterial exposure, the experiments showed. Nor did inflammatory chemicals called interleukins, or white cells, which are activated during allergic reactions and are also known to be elevated in skin diseases and even in certain neurologic disorders. "When we started the study, it was unclear whether the itch was a result of inflammation or not," Deng said. "We show that these things can be decoupled, that you don't necessarily have to have inflammation for the microbe to cause itch, but that the itch exacerbates inflammation on the skin." Interrupting the itch-scratch cycle Because PAR1 -- the protein activated by S. aureus -- is involved in blood-clotting, researchers wanted to see whether an already approved anticlotting drug that blocks PAR1 would stop itch. It did. The itchy mice whose skin was exposed to S. aureus experienced rapid improvement when treated with the drug. Their desire to scratch diminished dramatically, as did the skin damage caused by scratching. Moreover, once treated with PAR1 blockers, the mice no longer experienced abnormal itch in response to innocuous stimuli. The PAR1 blocker is already used in humans to prevent blood clots and could be repurposed as anti-itch medication. For example, the researchers noted, the active ingredient in the medicine could become the basis for anti-itch topical creams. One immediate question that the researchers plan to explore in future work is whether other microbes besides S. aureus can trigger itch. "We know that many microbes, including fungi, viruses, and bacteria, are accompanied by itch but how they cause itch is not clear," Chiu said. Beyond that, the findings raise a broader question: Why would a microbe cause itch? Evolutionarily speaking, what's in it for the bacterium? One possibility, the researchers said, is that pathogens may hijack itch and other neural reflexes to their advantage. For example, previous research has shown that the TB bacterium directly activates vagal neurons to cause cough, which might enable it to spread more easily from one host to another. "It's a speculation at this point, but the itch-scratch cycle could benefit the microbes and enable their spread to distant body sites and to uninfected hosts," Deng said. "Why do we itch and scratch? Does it help us, or does it help the microbe? That's something that we could follow up on in the future." The work was funded by the National Institutes of Health (grants R01AI168005, R01AI153185, R01NS065926, R01NS102161, R01NS111929, R37AI052453, R01AR076082, U01AI152038, UM1AI151958, R01AI153185, R01JL160582, F32AI172080, T32AI049928, 1R21AG075419), Food Allergy Science Initiative (FASI), Burroughs Wellcome Fund, Drako Family Fund, Jackson-Wijaya Research Fund, Canadian Institutes of Health Research (CIHR) (grants 376560 and 469411), and ANR-PARCURE (PRCE-CE18, 2020). Chiu serves on the scientific advisory board of GSK Pharmaceuticals. Provisional patent application Serial No. 63/438,668, in which some coauthors are listed as inventors, was filed based on these findings.

CINCINNATI, Ohio, June 14, 2023 (GLOBE NEWSWIRE) -- Blue Water Biotech, Inc. (Nasdaq: BWV), a biotechnology company spanning multiple sectors, today announced the signing of an Asset Purchase Agreement (the “Agreement”) with WraSer, LLC and Xspire Pharma, LLC (together, “WraSer”) for the purchase of six FDA-approved assets across multiple indications, including cardiology, otic infections, and pain management (the “WraSer Assets”). Under the terms of the Agreement, Blue Water Biotech, Inc. (“Blue Water” or the “Company”) will purchase the WraSer Assets for a total of $8.5 million cash consideration payable in three tranches: at signing, upon closing and within one year of closing. In addition, WraSer shall receive 1.0 million restricted shares of Blue Water’s common stock on the closing date. The closing of the transaction is subject to certain customary closing conditions, as well as WraSer’s delivery of 2022 audited financial statements for WraSer’s parent company, Legacy-Xspire Holdings, LLC, to Blue Water. Blue Water will assume management of the WraSer Assets during the audit period. The audited financial statements will be publicly filed promptly after completion of the audit. The assets under the Agreement include the patents related to ZONTIVITY®, a medication for the reduction of thrombotic cardiovascular events in patients with a history of myocardial infarction (“MI”) or with peripheral arterial disease (“PAD”), as well as additional products for the treatment of hypertension, otitis media, and pain management. ZONTIVITY®, a once daily pill approved in 2014, is a first-in-class protease activated receptor-1 inhibitor which mediates the cellular effects of thrombin to reduce the risk of major adverse cardiovascular events and major adverse limb events when used in addition to aspirin and/or clopidogrel in patients with PAD. In a pivotal Phase 3 clinical trial, ZONTIVITY® was shown to reduce the rate of a combined endpoint of cardiovascular death, MI, stroke, and urgent coronary revascularization, with 13,186 patients evaluated for safety and 2,187 patients treated with ZONTIVITY® for more than three years. This Agreement also includes the purchase of licenses to market and distribute two antibiotic otic products, OTOVEL® and CETRAXAL®, and their authorized generics currently marketed and distributed by WraSer. OTOVEL®, ciprofloxacin and fluocinolone acetonide, is used to treat acute otitis media (“AOM”) in children 6 months of age and older who have a tiny cylinder tube, known as a tympanostomy tube, inserted in their eardrum that prevents excess fluid in the middle ear. While tympanostomy tubes are typically placed as a result of multiple ear infections within a short timeframe, they do not prevent AOM, and one study found that over 80% of children had an episode of AOM within 18 months of tympanostomy tube placement. OTOVEL®, which contains an antibiotic to treat the infection and an anti-inflammatory to reduce pain and swelling, is the first and only antibiotic and steroid ear drop available in single use vials. In two clinical studies of pediatric patients with AOM with tympanostomy tubes, children who took OTOVEL® were clear of otorrhea in a median of 3.75 and 4.94 days, whereas those on antibiotic alone were not clear until a median of 7.69 and 6.83 days. The clinical studies also showed that the combination of medicines in OTOVEL® had similar side effects as the separate medicines. Blue Water is currently developing a vaccine to prevent acute otitis media regardless of tympanostomy tube insertion, and the Company intends to leverage this experience and expertise in the future commercialization and marketing for OTOVEL®. The second otic product included in the Agreement, CETRAXAL®, ciprofloxacin otic solution available in individual unit dosing vials, is used to treat otitis externa, more commonly called swimmer’s ear, in children with an intact tympanic membrane. Swimmer’s ear is characterized by redness or swelling, irritation, or infection of the outer ear canal and can be very painful if left untreated, primarily impacting children that spend a lot of time in the water. In addition to ZONTIVITY® and otic products, Blue Water will purchase the licensed rights to market and distribute CONJUPRI®, a calcium channel blocker that can be used alone or in combination with other antihypertensive agents for the treatment of hypertension to lower blood pressure. Lowering blood pressure reduces the risk of fatal and nonfatal cardiovascular events, primarily strokes and MIs. Additionally, Blue Water will purchase two pain medications, TREZIX™ (acetaminophen, caffeine, and dihydrocodeine bitartrate), for the treatment of moderate to moderately severe pain for which non-opioids are inadequate, and NALFON® (fenoprofen calcium), for relief of mild to moderate pain in adults, as well as relief of the signs and symptoms related to rheumatoid arthritis and osteoarthritis. “The team at WraSer has developed a strong pipeline across cardiology, otic products, as well as pain management, and we are thrilled to add these assets to our portfolio at Blue Water,” said Joseph Hernandez, Chairman and Chief Executive Officer of Blue Water. “In recent months, we have invested significant efforts to build our corporate infrastructure to support our transition into a commercial-stage company, including key hires with extensive experience in commercial operations. We are confident that bringing these assets into our portfolio will strengthen Blue Water’s financial position, allow us to advance our robust vaccine pipeline, and help millions of patients around the world receive proper access to critical treatments.” “Our products have been developed to provide patients with life changing, and potentially lifesaving, treatments, and we are happy to partner with Blue Water to continue this mission,” said Greg Stokes, Chief Executive Officer of WraSer. “Their management team has extensive experience with commercial products across a wide range of treatment areas, and we look forward to supporting them through this transition, as well as seeing the continued success of these products for patients in need.” Blue Water, with its recent purchase of ENTADFI® and through this acquisition, has established itself as a commercial-stage biopharmaceutical company. The Company intends to leverage its robust management and commercial team curated to maximize sales and success of all products acquired from the Agreement, as well as ENTADFI®, to further develop corporate infrastructure and to support and its extensive vaccine pipeline. About Blue Water Biotech Blue Water Biotech, Inc. is a biological and pharmaceutical technology company focused on developing and providing transformational therapies to address significant health challenges globally. Headquartered in Cincinnati, OH, the Company owns ENTADFI®, an FDA-approved, once daily pill that combines finasteride and tadalafil for the treatment of benign prostatic hyperplasia. This combination allows men to receive treatment for their symptoms of benign prostatic hyperplasia without the negative sexual side effects typically seen in patients on finasteride alone. The Company also has a robust vaccine pipeline. Blue Water holds the rights to proprietary technology developed at the University of Oxford, Cincinnati Children’s Hospital Medical Center, St. Jude Children’s Hospital, and The University of Texas Health Science Center at San Antonio. Blue Water is developing a Streptococcus pneumoniae vaccine candidate, designed to specifically prevent highly infectious middle ear infections, known as AOM, in children, and prevention of pneumonia in the elderly. The Company is also developing a universal flu vaccine that will provide protection from all virulent strains in addition to licensing a novel norovirus S&P nanoparticle versatile virus-like particle vaccine platform from Cincinnati Children’s to develop vaccines for multiple infectious diseases, including Marburg and monkeypox, among others. Additionally, the Company is developing a Chlamydia vaccine candidate with UT Health Science Center San Antonio to prevent infection and reduce the need for antibiotic treatment associated with contracting Chlamydia disease. For more information about Blue Water, visit www.bwbioinc.com. Forward-Looking Statements Certain statements in this press release are forward-looking within the meaning of the Private Securities Litigation Reform Act of 1995. These statements may be identified by the use of forward-looking words such as “anticipate,” “believe,” “forecast,” “estimate,” “expect,” and “intend,” among others. These forward-looking statements are based on Blue Water’s current expectations and actual results could differ materially. There are a number of factors that could cause actual events to differ materially from those indicated by such forward-looking statements. These factors include, but are not limited to, risks related to Blue Water’s ability to realize the benefits of its acquisitions of ENTADFI®, ZONTIVITY®, OTOVEL®, CETRAXAL®, CONJUPRI®, TREZIX™ and NALFON®, risks related to Blue Water’s ability to expand its business scope, commercialize ENTADFI® and integrate the assets and commercial operations acquired from WraSer into Blue Water’s business, risks related to the development of Blue Water’s vaccine candidates; the failure to obtain FDA clearances or approvals and noncompliance with FDA regulations; delays and uncertainties caused by the global COVID-19 pandemic; risks related to the timing and progress of clinical development of our product candidates; our need for additional financing; uncertainties of patent protection and litigation; uncertainties of government or third party payor reimbursement; limited research and development efforts and dependence upon third parties; and substantial competition. As with any vaccine under development, there are significant risks in the development, regulatory approval and commercialization of new products. Blue Water does not undertake an obligation to update or revise any forward-looking statement. Investors should read the risk factors set forth in Blue Water’s Annual Report on Form 10-K, filed with the Securities and Exchange Commission (the “SEC”) on March 9, 2023 and periodic reports filed with the SEC on or after the date thereof. All of Blue Water’s forward-looking statements are expressly qualified by all such risk factors and other cautionary statements. The information set forth herein speaks only as of the date thereof. Media Contact Information:Blue Water Media RelationsTelephone: (646) 942-5591 Email: Nic.Johnson@russopartnersllc.com Investor Contact Information:Blue Water Investor RelationsEmail: investors@bwbioinc.com A photo accompanying this announcement is available at https://www.globenewswire.com/NewsRoom/AttachmentNg/d66c95d4-fd36-4f08-9971-f362edf2a98a