Application of PI3K inhibitor in preparation of medicine for resisting high-toxicity klebsiella pneumoniae infection

Abstract

The invention relates to the technical field of biological medicines, and particularly discloses application of a PI3K inhibitor in preparation of a medicine for resisting high-toxicity klebsiella pneumoniae infection. According to the application disclosed by the invention, the high-toxicity klebsiella pneumoniae can invade and survive in the macrophages, while the PI3K inhibitor Ly294002 can aggravate the death of the macrophages caused by the infection, and the intracellular environment of the high-toxicity klebsiella pneumoniae is destroyed in the process, so that the effect of inhibiting the survival and proliferation of the high-toxicity klebsiella pneumoniae is achieved. The invention provides a novel anti-infection treatment strategy which does not depend on direct sterilization of antibiotics, and the strategy not only provides a new way for treatment of high-toxicity klebsiella pneumoniae infection with antibiotic resistance, but also contributes to reduction of clinical antibiotic selection pressure, so that development of drug resistance is indirectly delayed.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the application of PI3K inhibitors in the preparation of drugs against highly virulent Klebsiella pneumoniae infection. Background Technology

[0002] Hypervirulent Klebsiella pneumoniae Klebsiella pneumoniae Klebsiella pneumoniae (HVKP) is an important clinical subtype of Klebsiella pneumoniae, characterized by its strong pathogenicity and invasiveness, posing a significant public health threat. Currently, severe infections caused by highly virulent Klebsiella pneumoniae and its multidrug-resistant strains are primarily treated with various antibiotics. However, the widespread use of antibiotics continues to drive the evolution of this pathogen towards higher drug resistance and greater virulence, leading to a sustained increase in the incidence of multidrug-resistant highly virulent Klebsiella pneumoniae, placing a heavy burden on public health. Therefore, it is urgent to develop antibiotic-free prevention and control strategies to address multidrug-resistant highly virulent Klebsiella pneumoniae infections. Summary of the Invention

[0003] To address antibiotic resistance in highly virulent Klebsiella pneumoniae and explore novel antibiotic-free treatment pathways, this invention provides the application of PI3K inhibitors in the preparation of drugs against highly virulent Klebsiella pneumoniae infections. The PI3K inhibitor Ly294002 promotes macrophage death induced by highly virulent Klebsiella pneumoniae infection, thereby disrupting the bacterium's intracellular survival environment and ultimately inhibiting its intracellular survival and proliferation without increasing Klebsiella pneumoniae resistance. This provides a new strategy for non-antibiotic treatment of highly virulent Klebsiella pneumoniae infections.

[0004] Based on the above findings, the present invention provides the following technical solution: This invention provides the application of PI3K inhibitors in the preparation of drugs against highly virulent Klebsiella pneumoniae infection.

[0005] In some implementations, the PI3K inhibitor is Ly294002 or a pharmaceutically acceptable salt thereof or a derivative thereof.

[0006] In some implementations, the pharmaceutically acceptable salt is a sulfate or hydrochloride.

[0007] In some embodiments, the antiviral Klebsiella pneumoniae infection drug is a drug used to treat diseases caused by highly virulent Klebsiella pneumoniae infection.

[0008] In some implementations, the disease is one or more of the following: primary liver abscess, pneumonia, endophthalmitis, necrotizing fasciitis, purulent meningitis, and sepsis.

[0009] In some implementations, highly virulent Klebsiella pneumoniae infections can be distinguished etiologically by the following characteristics: (1) The highly virulent Klebsiella pneumoniae exhibits a high mucus phenotype; (2) The virulence gene of the highly virulent Klebsiella pneumoniae was positive, and the virulence gene includes: rmpA , rmpA2 , iucA , iroB , magA One or more genes.

[0010] In some implementations, highly virulent Klebsiella pneumoniae infections are clinically differentiated by one or more of the following characteristics: (1) Community-acquired, invasive infections; (2) It can cause metastatic foci of infection in healthy individuals without a clear underlying hepatobiliary disease; (3) Formation of a primary liver abscess caused by only one pathogen, Klebsiella pneumoniae, accompanied by one or more extrahepatic complications, such as endophthalmitis, meningitis, or necrotizing fasciitis.

[0011] In some implementations, the drug also includes a pharmaceutically acceptable carrier of a PI3K inhibitor.

[0012] In some implementations, the drug uses a PI3K inhibitor as the sole active ingredient against highly virulent Klebsiella pneumoniae infection.

[0013] In some embodiments, the dosage form of the drug is an injection; the injection is an injection solution and / or a lyophilized powder for injection.

[0014] Compared with the prior art, the present invention has at least the following beneficial effects: This invention proposes that highly virulent Klebsiella pneumoniae can invade and survive within macrophages, and Ly294002 can further promote infection-induced macrophage death, thereby disrupting the intracellular host environment upon which the bacteria depend for survival, ultimately inhibiting the bacteria's own survival and proliferation. LDH assay results showed that under conditions of highly virulent Klebsiella pneumoniae infection, the amount of LDH released in the experimental group treated with Ly294002 was significantly higher than that in the control group treated with DMSO, confirming that Ly294002 can exacerbate infection-induced macrophage death. Because Ly294002 exerts its antibacterial effect by affecting host cells, it is less likely to induce drug resistance in highly virulent Klebsiella pneumoniae compared to traditional antibiotic treatment. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 The study investigated the survival and death induction of highly virulent Klebsiella pneumoniae in macrophages and its effect on AKT phosphorylation. Where: a represents the survival of highly virulent Klebsiella pneumoniae in macrophages; the x-axis represents treatment time; and the y-axis represents the fold change in the number of surviving highly virulent Klebsiella pneumoniae bacteria in cells relative to the initial number after 1 hour of infection. This indicates that p < 0.01. This indicates that p < 0.001. This indicates p < 0.0001; b represents the ability of highly virulent Klebsiella pneumoniae infection to induce macrophage death, where: CK group represents PBS treatment, Kp group represents Klebsiella pneumoniae bacterial suspension treatment; the horizontal axis represents treatment time; the vertical axis represents LDH release. This indicates that p < 0.01. This indicates that p < 0.001. p < 0.0001; c represents the effect of Western blotting on the PI3K / AKT signaling pathway by highly virulent Klebsiella pneumoniae. The CK group was treated with PBS, and the Kp group was treated with highly virulent Klebsiella pneumoniae culture for 0.5, 1, 1.5, 5, 10, 12 and 24 hours.

[0017] Figure 2 The study aimed to promote the death of macrophages infected with highly virulent Klebsiella pneumoniae using Ly294002. In this study, a represents the effect of Ly294002 on macrophage death induced by highly virulent Klebsiella pneumoniae; the DMSO group represented untreated macrophages, and the Ly294002 group represented macrophages treated with 10 μg / mL Ly294002. The x-axis represents the amount of Kp without highly virulent Klebsiella pneumoniae treatment, and the y-axis represents the amount of LDH released. This indicates that p < 0.01. This indicates that p < 0.001. p < 0.0001; b represents the effect of Ly294002 on the survival of highly virulent Klebsiella pneumoniae in macrophages; where: DMSO group represents treatment with an equal volume of DMSO, Ly294002 group represents treatment with 10 μg / mL Ly294002; the x-axis represents treatment time; the y-axis represents the fold change in the number of highly virulent Klebsiella pneumoniae surviving in cells relative to the initial number in cells 1 hour after infection. This indicates that p < 0.01. This indicates that p < 0.001. This means p < 0.0001.

[0018] Figure 3 To investigate the effect of cell death on the survival of Klebsiella pneumoniae in macrophages; where: a) treatment with 10 μM Z-VAD-fmk on Klebsiella pneumoniae-infected macrophages; b) the effect of Z-VAD-fmk treatment for 1 hour or 6 hours on the survival of Klebsiella pneumoniae in macrophages. This indicates that p < 0.01. This indicates that p < 0.001. This means p < 0.0001.

[0019] Figure 4 The effect of Ly294002 on the lethality of mice infected with highly virulent Klebsiella pneumoniae was investigated. The WT+DMSO group consisted of male C57BL / 6J mice infected with highly virulent Klebsiella pneumoniae and treated with DMSO, while the WT+Ly294002 group consisted of male C57BL / 6J mice infected with highly virulent Klebsiella pneumoniae and treated with Ly294002. The x-axis represents treatment time, and the y-axis represents survival probability. This indicates that p < 0.01. This indicates that p < 0.001. This means p < 0.0001. Detailed Implementation

[0020] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below in conjunction with the embodiments of this invention. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0021] Ly294002 is a widely used and highly effective cell-permeable small molecule inhibitor of synthetic PI3K. It competitively binds to the ATP-binding pocket of PI3K, reversibly inhibiting the catalytic activity of the enzyme and thus effectively blocking downstream PI3K / Akt signaling pathways.

[0022] This invention provides the application of PI3K inhibitors in the preparation of drugs against highly virulent Klebsiella pneumoniae infection. Highly virulent Klebsiella pneumoniae is a highly invasive Gram-negative bacillus that can cause severe infection not only in immunocompromised individuals but also pose a threat to healthy individuals. Experiments show that the PI3K inhibitor Ly294002 can promote macrophage death after infection with highly virulent Klebsiella pneumoniae, thereby inhibiting the survival of this bacterium within host cells.

[0023] In some embodiments, the PI3K inhibitor is Ly294002 or a pharmaceutically acceptable salt thereof or a derivative thereof. Ly294002, whose Chinese name is 2-morpholino-8-phenylchromone, has the following structural formula:

[0024] Its pharmaceutically acceptable salt is an ionic compound that meets pharmaceutical standards and is formed by the reaction of Ly294002 molecules with acids or bases through salt formation; its derivatives are formed by the chemical or physical modification of Ly294002 molecules.

[0025] In some implementations, the pharmaceutically acceptable salt is a sulfate or hydrochloride.

[0026] In some embodiments, the antiviral drug for Klebsiella pneumoniae infection is a drug used to treat diseases caused by infection with highly virulent Klebsiella pneumoniae. This drug has the following therapeutic effects, including: it can effectively kill pathogens in the blood and tissues, effectively control primary and metastatic foci of infection, and achieve rapid clearance of the pathogens; it can block the further spread of bacteria to other organs such as the eyes and brain; it can promptly terminate the course of sepsis, curb the deterioration of the condition, and prevent the development of septic shock and multiple organ failure; at the same time, it can completely eradicate the infection foci and effectively prevent recurrence of infection.

[0027] In some implementations, the disease is one or more of the following: primary liver abscess, pneumonia, endophthalmitis, necrotizing fasciitis, purulent meningitis, and sepsis. After highly virulent Klebsiella pneumoniae proliferates extensively within the primary focus of infection, it uses its high mucoid phenotype and virulence factors to disrupt local tissue barriers, directly invading or indirectly entering the vascular system via lymphatic drainage, thereby causing bacteremia. It then disseminates via the bloodstream to richly vascularized organs such as the liver, spleen, eyes, central nervous system, lungs, and joints, forming new purulent foci of infection.

[0028] In some implementations, highly virulent Klebsiella pneumoniae infection is characterized etiologically by: on the one hand, a high mucus phenotype, and on the other hand... rmpA , rmpA2 , iucA , iroB , magA The virulence gene test was positive. The high mucus phenotype is the most obvious external marker of highly virulent Klebsiella pneumoniae and is the physical basis for its systemic dissemination and resistance to immune clearance. rmpA and rmpA2 It is responsible for regulating the expression of genes related to capsular polysaccharide synthesis, which in turn leads to a high-mucus phenotype; iucA , iroB Its function is to help highly virulent Klebsiella pneumoniae obtain iron ions for its own proliferation; magA It is a K1 capsule serotype-specific gene, and its high expression means that the strain belongs to the high-risk K1 type.

[0029] In some implementations, highly virulent Klebsiella pneumoniae infection presents with the following clinical characteristics: First, it is characterized by community-acquired and invasive infection, unlike classic Klebsiella pneumoniae which mainly infects immunocompromised individuals in hospitals or medical institutions. Highly virulent Klebsiella pneumoniae can infect relatively healthy individuals without clear immunodeficiency in the community environment. Second, it can induce metastatic infection foci in healthy individuals without underlying hepatobiliary diseases. Conventional bacterial liver abscesses are more common in people with biliary tract diseases, diabetes, or immunocompromised individuals, while highly virulent Klebsiella pneumoniae can induce liver abscesses in healthy individuals without primary hepatobiliary diseases and further form new infection foci in other distal sites of the body. Third, it can cause primary liver abscesses caused by a single pathogen, highly virulent Klebsiella pneumoniae, and is often accompanied by one or more extrahepatic complications, such as endophthalmitis, meningitis, or necrotizing fasciitis.

[0030] In some embodiments, the drug also includes a pharmaceutically acceptable carrier for the PI3K inhibitor. The carrier is capable of mixing, binding, or dissolving with the active ingredient, the PI3K inhibitor, during formulation, thereby facilitating the formation of a specific dosage form that is safe, stable, effective, and suitable for administration.

[0031] In some embodiments, the drug uses a PI3K inhibitor as the sole active ingredient against highly virulent Klebsiella pneumoniae infection. Specifically, the drug does not contain conventional antibiotics or other antibacterial agents, and its ability to combat virulent Klebsiella pneumoniae infection relies entirely on the regulatory effect of the PI3K inhibitor on host cells, rather than directly killing the bacteria.

[0032] In some embodiments, the drug is in the form of an injection; the injection is an injection solution and / or a lyophilized powder for injection. Compared with conventional drug dosage forms, the core advantage of injection dosage forms lies in their ability to perfectly meet the treatment needs of acute and critical illnesses such as highly virulent Klebsiella pneumoniae infection, and they have the characteristics of high bioavailability, rapid onset of action, and no limitation by the patient's state of consciousness or gastrointestinal function.

[0033] Ly294002 and Z-VAD-fmk used in the following examples were purchased from MCE, and phorbol ester was purchased from Sigma.

[0034] Example 1: Highly virulent Klebsiella pneumoniae can survive in macrophages. PMA was dissolved in DMSO to prepare a stock solution with a concentration of 100 ng / μL, which was then stored at -20°C. THP-1 cells were subsequently differentiated in a medium containing 25 ng / mL PMA for 48 hours, resulting in a cell concentration of approximately 1 × 10⁻⁶ cells. 6 / mL, after differentiation, replace with complete medium without PMA (1640 basal medium + 5% FBS + 100 μg / mL gentamicin), and continue culturing for 12 hours. Simultaneously, culture highly virulent Klebsiella pneumoniae in the logarithmic phase, centrifuge at 6000 rpm for 10 minutes, and adjust the bacterial count to OD using PBS. 600 Set the value to 0.5 and use it as a backup. At this point, the bacterial concentration is approximately 1 × 10⁻⁵. 8 CFU / mL. Differentiated THP-1 macrophages were infected at a bacterial-to-cell ratio of approximately 50:1 for 1 hour, followed by washing with PBS to remove unphagocytic highly virulent Klebsiella pneumoniae. Complete culture medium (1640 basal medium + 5% FBS + 100 μg / mL gentamicin) was added to the macrophages, and they were cultured for another 6 hours. Macrophages were harvested after 1 hour and 6 hours of culture, washed three times with PBS, and lysed with 0.5% saponin. The cells were then serially diluted and plated for counting. Plate counting results showed that the number of highly virulent Klebsiella pneumoniae infecting macrophages significantly increased after 6 hours of culture compared to 1 hour, indicating that highly virulent Klebsiella pneumoniae can colonize and survive within macrophages. Figure 1 a).

[0035] Example 2: Infection with highly virulent Klebsiella pneumoniae can induce macrophage death. THP-1 cells were differentiated in a medium containing 25 ng / mL PMA for 48 hours, resulting in a cell concentration of approximately 1 × 10⁻⁶ cells. 6 / mL, after differentiation, replace with complete medium without PMA, and continue culturing for 12 hours. Meanwhile, highly virulent Klebsiella pneumoniae cultured to the logarithmic growth phase was centrifuged at 6000 rpm for 10 minutes, and the bacterial count was adjusted to OD using PBS. 600 Set the value to 0.5 and use it as a backup. At this point, the bacterial concentration is approximately 1 × 10⁻⁵. 8 CFU / mL. Differentiated THP-1 macrophages were randomly divided into two groups: (1) CK group: treated with 100 μL PBS for 1 hour; (2) Kp group: 100 μL of highly virulent Klebsiella pneumoniae bacterial solution was used for infection for 1 hour.

[0036] One hour after infection, PBS was removed from the CK group, while the Kp group was washed with PBS to remove unphagocytic highly virulent Klebsiella pneumoniae. Both groups were then added to complete culture medium and cultured for another 6 hours. Finally, the supernatants from 1 and 6 hours of culture were collected, and cell death was detected using an LDH assay kit. The results showed that after 6 hours of culture, the LDH release in the Kp group was significantly higher than that in the CK group, indicating that infection with highly virulent Klebsiella pneumoniae can induce macrophage death. Figure 1 b).

[0037] Example 3: Effects of highly virulent Klebsiella pneumoniae on the PI3K / AKT signaling pathway THP-1 cells were differentiated with phorbol ester at a concentration of 25 ng / mL for 48 hours, resulting in a cell concentration of approximately 1 × 10⁻⁶. 6 / mL, after differentiation, replace with normal medium without PMA, and continue culturing for 12 hours. Meanwhile, highly virulent Klebsiella pneumoniae cultured to the logarithmic growth phase was centrifuged at 6000 rpm for 10 minutes, and the bacterial count was adjusted to OD using PBS. 600 Set the value to 0.5 and use it as a backup. At this point, the bacterial concentration is approximately 1 × 10⁻⁵. 8 CFU / mL. Differentiated THP-1 macrophages were randomly divided into two groups: (1) CK group: 100 μL PBS for 1 hour; (2) Kp group: 100 μL of highly virulent Klebsiella pneumoniae bacterial solution was used for infection for 1 hour.

[0038] One hour after infection, PBS was removed from the CK group, while the Kp group was washed with PBS to remove unphagocytic highly virulent Klebsiella pneumoniae. The cells were then cultured in complete medium for 0.5, 1, 1.5, 5, 10, 12, and 24 hours, respectively. The supernatant was discarded, and the cells were washed once with PBS. 100 μL of M-per cell lysis buffer containing a cocktail protease inhibitor was added to lyse the cells and extract proteins. Western blotting was used to detect the phosphorylation level of AKT. The results showed that the phosphorylation level of AKT began to decrease significantly five hours after infection with highly virulent Klebsiella pneumoniae. Figure 1 c), this result indicates that infection with highly virulent Klebsiella pneumoniae can inhibit the activation of the AKT signaling pathway.

[0039] Example 4: Ly294002 promotes macrophage death induced by highly virulent Klebsiella pneumoniae. Ly294002 was dissolved in DMSO to prepare a stock solution with a concentration of 10 μg / μL, which was then stored at -20℃. Subsequently, THP-1 cells were differentiated with 25 ng / mL PMA for 48 hours, resulting in a cell concentration of approximately 1×10⁻⁶ cells. 6 / mL, after differentiation, replace with normal medium without PMA, and continue culturing for 12 hours. Meanwhile, highly virulent Klebsiella pneumoniae cultured to the logarithmic growth phase was centrifuged at 6000 rpm for 10 minutes, and the bacterial count was adjusted to OD using PBS. 600 Set the value to 0.5 and use it as a backup. At this point, the bacterial concentration is approximately 1 × 10⁻⁵. 8 CFU / mL. The experimental setup was as follows: (1) -Kp+DMSO group: 100 μL PBS+1 μL DMSO; (2) Kp+Ly294002 group: 100 μL PBS + 1 μL 10 μg / μL Ly294002, so that the final concentration of Ly294002 is 10 μg / mL; (3) +Kp+DMSO group: 100 μL of highly virulent Klebsiella pneumoniae infection + 1 μL of DMSO; (4) +Kp+Ly294002 group: 100 μL of highly virulent Klebsiella pneumoniae infection + 1 μL of 10 μg / μL Ly294002, so that the final concentration of Ly294002 is 10 μg / mL.

[0040] For the infected groups (+Kp+DMSO group, +Kp+Ly294002 group), highly virulent Klebsiella pneumoniae and macrophages were co-incubated at a ratio of approximately 50:1 for 1 hour, followed by washing with PBS to remove unphagocytosed bacteria. For the uninfected groups (-Kp+DMSO group, -Kp+Ly294002 group), an equal volume of PBS was added. All groups were then replaced with complete culture medium, and 1 μL of 10 μg / μL Ly294002 or an equal volume of DMSO solution was added according to the group to bring the final Ly294002 concentration to 10 μg / mL, and cultured for another 6 hours. After culture, the supernatant was collected, and cell death was detected using an LDH detection kit. The results showed no significant difference in LDH release between the -Kp+DMSO group and the -Kp+Ly294002 group, but the LDH release in the +Kp+Ly294002 group was significantly higher than that in the +Kp+DMSO group. Figure 2 (a) This result demonstrates that Ly294002 can promote macrophage death induced by highly virulent Klebsiella pneumoniae.

[0041] Example 5: The Ly294002 group significantly inhibited the survival of highly virulent Klebsiella pneumoniae in macrophages. Ly294002 was dissolved in DMSO to prepare a stock solution with a concentration of 10 μg / μL, which was then stored at -20℃. Subsequently, THP-1 cells were differentiated with 25 ng / mL PMA for 48 hours, resulting in a cell concentration of approximately 1×10⁻⁶ cells. 6 / mL, after differentiation, replace with normal medium without PMA, and continue culturing for 12 hours; at the same time, culture highly virulent Klebsiella pneumoniae in the logarithmic phase, centrifuge at 6000 rpm for 10 minutes, and adjust the bacterial count to OD using PBS. 600 The value is 0.5 for later use; at this point, the bacterial concentration is approximately 1 × 10⁻⁵. 8 CFU / mL. Differentiated THP-1 macrophages were infected at a ratio of approximately 50:1 of highly virulent Klebsiella pneumoniae to cells for 1 hour. Cells were then washed with PBS to remove unphagocytosed highly virulent Klebsiella pneumoniae. After removing the PBS, complete culture medium containing 10 μg / mL Ly294002 or an equal volume of DMSO was added to the cells. Cells were cultured for 1 hour or 6 hours, washed three times with PBS, and lysed with 0.5% saponin. The cells were then serially diluted and plated for counting. Plate counting results showed that, compared to the DMSO group, the Ly294002 group had no significant effect on the survival of highly virulent Klebsiella pneumoniae in macrophages after 1 hour of culture; however, after 6 hours of culture, the Ly294002 group significantly inhibited the survival of highly virulent Klebsiella pneumoniae in macrophages compared to the DMSO group. Figure 2 b).

[0042] Example 6: Effect of inhibiting macrophage death on the survival of Klebsiella pneumoniae Z-VAD-fmk was dissolved in DMSO and prepared as a 10 mM stock solution, which was stored at -20°C. THP-1 cells were then differentiated with 25 ng / mL phorbol ester for 48 hours. Differentiated THP-1 macrophages were then infected for 1 hour at a bacterial-to-cell ratio of approximately 50:1. Unphagocytic highly virulent Klebsiella pneumoniae were washed away with PBS, and cells were cultured for 1 hour or 6 hours in complete medium containing either 10 μM pan-caspase inhibitor Z-VAD-fmk or an equal volume of DMSO. Cells after 1 and 6 hours of culture were washed three times with PBS, lysed with 0.5% saponin, serially diluted, plated, and counted. The results showed that Z-VAD-fmk had no significant effect on the survival of highly virulent Klebsiella pneumoniae in macrophages after 1 hour of culture. However, after 6 hours of culture, compared to the DMSO group, Z-VAD-fmk treatment promoted the survival of highly virulent Klebsiella pneumoniae in macrophages. Figure 3 b). Additionally, the supernatant from 6 hours of culture was used to detect cell death using an LDH detection kit. The results showed that the LDH release in the Z-VAD-fmk group was significantly lower than that in the DMSO group ( Figure 3 a).

[0043] Example 7: Effect of Ly294002 treatment on mortality rate of mice infected with highly virulent Klebsiella pneumoniae.

[0044] Highly virulent Klebsiella pneumoniae cultured to the logarithmic growth phase were centrifuged at 6000 rpm for 10 minutes, the supernatant was discarded, and the bacterial concentration was adjusted to 4 × 10⁻⁶ using PBS. 6 CFU / mL was kept on hand. Male C57BL / 6J mice aged 5-6 weeks were purchased from Jicui Pharmaceutical Co., Ltd. After anesthetizing with isoflurane, 30 μL of the prepared bacterial solution was instilled into the nasal cavity of each mouse. The infectious dose per mouse was 1×10⁻⁶. 5 CFU. Six hours after infection, mice were randomly divided into two groups: WT+DMSO and WT+Ly294002, with eight mice in each group. Mice in the WT+Ly294002 group were intraperitoneally injected with 200 μL of Ly294002 solution (5 mg / kg Ly294002, PBS as solvent), while mice in the WT+DMSO group were intraperitoneally injected with an equal volume of DMSO solution. The condition and survival of the mice were observed and recorded every 4 hours after administration. The results showed that Ly294002 treatment significantly improved the survival rate of mice infected with highly virulent Klebsiella pneumoniae. Figure 4 ).

[0045] In the description of this specification, the references to terms such as "one embodiment / mode," "some embodiments / modes," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment / mode or example is included in at least one embodiment / mode or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment / mode or example. Moreover, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments / modes or examples. Furthermore, without contradiction, those skilled in the art can combine and integrate the different embodiments / modes or examples described in this specification, as well as the features of different embodiments / modes or examples.

[0046] It should be noted that in this invention, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element. In this invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise expressly specified.

[0047] The above description is merely a specific embodiment of the present invention, enabling those skilled in the art to understand or implement the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the present invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features claimed herein.

Claims

1. Application of PI3K inhibitors in the preparation of drugs against highly virulent Klebsiella pneumoniae infection.

2. The application according to claim 1, characterized in that: The PI3K inhibitor is Ly294002 or a pharmaceutically acceptable salt thereof or a derivative thereof.

3. The application according to claim 2, characterized in that: The pharmaceutically acceptable salt is a sulfate or hydrochloride.

4. The application according to claim 1, characterized in that: The aforementioned antiviral Klebsiella pneumoniae infection drug is used to treat diseases caused by highly virulent Klebsiella pneumoniae infection.

5. The application according to claim 4, characterized in that: The disease is one or more of the following: primary liver abscess, pneumonia, endophthalmitis, necrotizing fasciitis, purulent meningitis, and sepsis.

6. The application according to claim 1, characterized in that: The highly virulent Klebsiella pneumoniae infection is distinguished etiologically by the following characteristics: (1) The highly virulent Klebsiella pneumoniae exhibits a high mucus phenotype; (2) The virulence gene of the highly virulent Klebsiella pneumoniae was detected positive, and the virulence gene includes rmpA , rmpA2 , iucA , iroB , magA One or more genes.

7. The application according to claim 1, characterized in that: The highly virulent Klebsiella pneumoniae infection can be distinguished clinically by one or more of the following characteristics: (1) Community-acquired, invasive infections; (2) It can cause metastatic foci of infection in healthy individuals without a clear underlying hepatobiliary disease; (3) The formation of a primary liver abscess caused by only one pathogen, Klebsiella pneumoniae, accompanied by one or more extrahepatic complications, such as endophthalmitis, meningitis, or necrotizing fasciitis.

8. The application according to claim 1, characterized in that: The drug also includes pharmaceutically acceptable carriers of PI3K inhibitors.

9. The application according to claim 1, characterized in that: The drug uses a PI3K inhibitor as its sole effective component against highly virulent Klebsiella pneumoniae infection.

10. The application according to claim 1, characterized in that: The dosage form of the drug is an injection; the injection is an injection solution and / or a lyophilized powder for injection.

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