Use of hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) in rare anemia

Abstract

The present application relates to use of a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) in rare anemia. Specifically disclosed in the present application is use of certain hypoxia-inducible factor-prolyl hydroxylase inhibitors (HIF-PHIs) in the treatment of anemia of myelodysplastic syndromes (MDS anemia), beta-thalassemia (β-thalassemia), and / or sickle cell disease (sickle cell anemia, SCD anemia).

Description

Application of hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) in rare anemias

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese invention patent application No. 202311391912.1, filed on October 24, 2023, entitled “Application of Hypoxia-Inducible Factor-Prolyl Hydroxylase Inhibitors (HIF-PHI) in Rare Anemias,” the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present application belongs to the medical field, and in particular, the present application relates to the use of certain hypoxia-inducible factor-prolyl hydroxylase inhibitors (HIF-PHI) in the treatment of rare anemias. Background Art

[0004] The primary function of human red blood cells is to absorb oxygen inhaled from the lungs onto their hemoglobin. This oxygen is then transported through the bloodstream to the body's tissues for cellular metabolism. Carbon dioxide produced by the cells is then absorbed by the hemoglobin in the red blood cells and circulated through the bloodstream to the lungs for excretion, completing a cycle of oxygen inhalation, oxygen transport, carbon dioxide transport, and excretion. Normal hemoglobin levels fluctuate with age, gender, and other factors. For example, normal hemoglobin levels for adult men range from 14 to 18 g / dL, while those for adult women range from 12 to 16 g / dL. Erythropoietin (EPO) is a key hormone that maintains the homeostasis of red blood cells and hemoglobin. In healthy individuals, hemoglobin levels are inversely correlated with endogenous EPO levels. Specifically, as hemoglobin decreases, endogenous EPO increases, promoting hemoglobin elevation and red blood cell growth to compensate for the decrease, thereby maintaining homeostasis.

[0005] Anemia can result if the absolute number of red blood cells decreases or if the hemoglobin level decreases. Anemia not only seriously affects the patient's physical health and quality of life, but can even threaten the patient's life if it is not corrected. There are many types of anemia, among which anemia caused by chronic kidney disease (CKD) is one of the most common anemias. There are also many more rare anemias that have long troubled patients and doctors, such as myelodysplastic syndrome anemia (MDS anemia), β-thalassemia (β-thalassemia), sickle cell disease (also known as sickle cell anemia, or SCD anemia). The clinical manifestations and biological mechanisms of common anemias such as chronic kidney disease anemia and rare anemias such as MDS anemia, β-thalassemia, and SCD anemia are completely different; the existing drug treatments for them are also not exactly the same, which is completely reasonable.

[0006] MDS anemia is a rare, heterogeneous, clonal stem cell disorder characterized by abnormal myeloid cell development, manifested by ineffective and dysplastic hematopoiesis, leading to mono- or polycytopenias and a high risk of transformation to acute myeloid leukemia. The global incidence of MDS anemia is approximately 2 to 12 per 100,000 patients. Eighty percent of patients develop the disease over the age of 60, with a higher prevalence in men than in women. Patients with MDS anemia can be divided into lower- and higher-risk groups. In addition to red blood cell transfusions, supportive treatment for anemia in MDS patients includes rhEPO-based drugs, which are the primary treatment. However, the response rate to rheopolysaccharide (rhEPO) for MDS anemia is very low.

[0007] β-thalassemia (abbreviated as β-thalassemia) is a common, inherited, chronic blood disorder. Its pathogenesis is due to mutations or deletions in the β-globin gene in hemoglobin (Hb), resulting in an imbalance in the number of globin α and β chains. This leads to defective or ineffective erythropoiesis, resulting in chronic anemia and complications. The highest incidence is in the Mediterranean region, Southeast Asia, and the Middle East. In my country, β-thalassemia patients are primarily distributed in southern regions such as Guangdong, Guangxi, Yunnan, Guizhou, Sichuan, Hong Kong, and Macau, with an average prevalence of approximately 2%. Transfusion-dependent thalassemia (TDT), also known as severe thalassemia, often develops in infancy and presents as a chronic, progressive hemolytic anemia, seriously threatening the patient's quality of life and even their life. If left untreated, severe β-thalassemia often leads to death before the age of five. Standardized lifelong blood transfusions and iron chelation therapy are the main treatments for severe β-thalassemia. Splenectomy is palliative therapy, and hematopoietic stem cell transplantation is the only cure for thalassemia. However, its application is limited by the lack of blood donations, high medical costs, and limited medical resources. Recently, gene therapy has also been used to treat β-thalassemia, but it is extremely expensive, has a very limited population, and its long-term safety is unknown. Recently, the TGF-β-targeting fusion protein red blood cell maturation agent Rotacept was approved for severe β-thalassemia, but many patients do not respond to this drug, and it has serious side effects such as joint pain and bone pain. Severe thalassemia is a rare disease in both my country and the United States.

[0008] SCD anemia is another serious, life-threatening, inherited chronic blood disorder, predominantly affecting people of African descent. It affects approximately 100,000 people in the United States and over 3 million worldwide. It is considered a rare disease in both my country and the United States. The disease is caused by a mutation in the sixth amino acid of the hemoglobin β-globin from the hydrophilic, negatively charged glutamic acid to the lipophilic valine. This mutation causes hemoglobin in the cytoplasm of red blood cells to physically aggregate after oxygen molecules bound to tissues are exchanged, leading to the collapse of the red blood cells into a sickle shape. Clinically, this manifests as severe hemolytic anemia, and the deformed red blood cells block capillaries, causing ischemia, pain, and multiple organ failure. Hydroxyurea (HU) is the primary drug treatment for SCD anemia, but HU itself is a chemotherapy drug with some bone marrow suppression side effects, and long-term use can lead to leukemia. In addition, a small molecule drug, voxelotor, recently received conditional approval from the US FDA but has yet to receive final formal approval. Its mechanism of action is to embed itself into oxygen-free hemoglobin cavities, thereby preventing hemoglobin aggregation. However, its clinical efficacy is limited and it does not improve patients' quality of life (patient reported outcomes (PRO)).

[0009] Generally speaking, there is currently no safe and effective treatment for the above-mentioned rare anemias, such as myelodysplastic syndrome anemia (MDS anemia), β-thalassemia (β-thalassemia), and sickle cell disease (SCD anemia), especially no safe and effective small molecule drugs.

[0010] Therefore, there is a great need for a safe and effective small molecule drug for the treatment of rare anemias, especially rare anemias such as MDS anemia, β-thalassemia, and SCD anemia.

[0011] Summary of the Invention

[0012] In order to find safe and effective small molecule drugs that can be used to treat rare anemias, especially rare anemias such as MDS anemia, β-thalassemia, and SCD anemia, the inventors of the present application have conducted extensive research and investigation.

[0013] As a treatment for common anemias, such as anemia associated with chronic kidney disease (CKD), oral hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHIs) are gradually being approved by some regulatory agencies. As a novel small molecule drug, HIF-PHIs treat CKD by inhibiting the enzyme hypoxia-inducible factor prolyl hydroxylase, thereby inhibiting the degradation of HIF-α. This allows HIF-α to bind to HIF-β, forming a transcription factor with transcriptional function called a HIF dimer. This in turn activates multiple downstream target genes, including increasing endogenous EPO production, resulting in a therapeutic effect similar to recombinant erythropoietin in treating CKD anemia. According to public reports, HIF-PHIs that have entered clinical trials for the treatment of anemia in CKD include roxadustat, daprodustat, vadadustat, molidustat, enarodustat, desidustat, HIF117 (SSS17), HEC53856, DDO-3055, and FG-2216. Even so, not every HIF-PHI can truly safely and effectively treat anemia in CKD. For example, to date, only daprodustat has achieved success in a pivotal Phase III clinical trial in the United States for the treatment of anemia in patients with chronic kidney disease on dialysis, and its New Drug Application (NDA) has been approved by the FDA. Daprodustat has not achieved success in a pivotal Phase III clinical trial in the treatment of anemia in patients with non-dialysis chronic kidney disease, and its NDA has not been approved by the FDA. Roxadustat has also failed in pivotal Phase III clinical trials in both dialysis and non-dialysis patients with anemia in the United States, and its NDA has ultimately not been approved by the FDA. Therefore, even though many of the molecules listed above are considered HIF-PHIs, it is not obvious whether they can be proven safe and effective in treating anemia in chronic kidney disease.

[0014] Because HIF-PHIs stabilize HIF, activate EPO, and subsequently elevate EPO levels, they were once considered ideal alternatives to injectable recombinant erythropoietin (rEPO) for treating anemia in chronic kidney disease. For this reason, all publicly reported HIF-PHIs are being developed for the treatment of anemia in chronic kidney disease. Only roxadustat was tested for the treatment of anemia in MDS, but its efficacy in a Phase 3 clinical trial failed after failing to demonstrate significant difference compared to placebo. See the May 5, 2023 news release: https: / / www.biospace.com / article / releases / fibrogen-announces-results-for-matterhorn-a-phase-3-clinical-study-of-roxadustat-for-the-treatment-of-anemia-in-patients-with-myelodysplastic-syndromes-mds-study-did-not-meet-primary-endpoint / . Other HIF-PHIs are rarely developed for rare anemias, and the reason is easy to understand because the biological mechanisms of rare anemias such as MDS anemia, β-thalassemia, and SCD anemia are very different from those of chronic kidney disease anemia.

[0015] The failure of some of the aforementioned HIF-PHIs in pivotal Phase III clinical trials and NDAs for anemia in chronic kidney disease in the United States, as well as the success of dapoxetine in a pivotal Phase III clinical trial and FDA approval in the United States for anemia in chronic kidney disease with dialysis, demonstrate that while all hypoxia-inducible factor-proline hydroxylase inhibitors appear to have similar mechanisms of action, their safety and efficacy profiles in the most common anemias of chronic kidney disease vary significantly. This means that the safety and efficacy of these HIF-PHIs in anemia of chronic kidney disease are not readily apparent, let alone in rare anemias such as myelodysplastic syndrome (MDS), β-thalassemia, and sickle cell disease (SCD). Furthermore, the failure of roxadustat in a pivotal Phase III clinical trial for anemia in MDS in 2023 has made the application of various HIF-PHIs in rare anemias such as MDS, β-thalassemia, and SCD even more unpredictable.

[0016] However, unexpectedly, after in-depth research, the inventors of the present application found that compounds containing indoleazine derivatives (such as the compound synthesized in Example 24 of WO 2018205928) can effectively increase red blood cell count, hemoglobin content, and hematocrit in MDS anemia model mice, β-thalassemia model mice, and SCD anemia model mice, and improve MDS anemia, β-thalassemia, and SCD anemia; further unexpectedly, the above-mentioned indoleazine derivatives showed surprising effectiveness in MDS anemia model mice, β-thalassemia model mice, and sickle cell disease (sickle type) SCD anemia model mice, respectively; therefore, they have the effect of treating MDS anemia, β-thalassemia, and SCD anemia.

[0017] In addition, after further in-depth research, the inventors of the present application found that HIF-PHI molecules, which are hypoxia-inducible factor-proline hydroxylase inhibitors other than roxadustat, may have the function of treating MDS anemia or SCD anemia; and HIF-PHI molecules, which are hypoxia-inducible factor-proline hydroxylase inhibitors including roxadustat, may have the function of treating β-thalassemia.

[0018] Therefore, a first aspect of the present application provides use of a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) in the manufacture of a medicament for treating rare anemias.

[0019] A second aspect of the present application provides the use of a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) in the manufacture of a medicament for increasing red blood cell count, hemoglobin content and / or hematocrit in rare anemias.

[0020] The third aspect of the present application provides a method for treating rare anemias, comprising: administering a therapeutically effective amount of a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) to a patient in need of treatment.

[0021] The fourth aspect of the present application provides a method for increasing the red blood cell count, hemoglobin content and / or hematocrit of rare anemias, comprising: administering a therapeutically effective amount of a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) to a patient in need of treatment.

[0022] The fifth aspect of the present application provides a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) for treating rare anemias.

[0023] The sixth aspect of the present application provides the use of hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) for treating rare anemias. DETAILED DESCRIPTION

[0024] 1. Hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI)

[0025] In various aspects and various embodiments of the present application, the hypoxia-inducible factor-proline hydroxylase inhibitor can be various hypoxia-inducible factor-proline hydroxylase inhibitor compounds known in the art.

[0026] In some preferred embodiments of the present application, the hypoxia-inducible factor-proline hydroxylase inhibitor can be a HIF-PHI compound or a pharmaceutically acceptable salt thereof that has been approved for clinical use or is undergoing clinical trials. For example, the HIF-PHI can be selected from the group consisting of: Roxadustat, Daprodustat, Vadadustat, Molidustat, Enarodustat, Desidustat, HIF117 (SSS17), HEC53856, and DDO-3055; or the HIF-PHI can be selected from the group consisting of: Daprodustat, Vadadustat, Molidustat, Enarodustat, Desidustat, HIF117 (SSS17), HEC53856, and DDO-3055.

[0027] In some embodiments, the hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) is roxadustat or a pharmaceutically acceptable salt thereof.

[0028] In some embodiments, the hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) is Daprodustat or a pharmaceutically acceptable salt thereof.

[0029] In some embodiments, the hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) is Vadadustat or a pharmaceutically acceptable salt thereof.

[0030] In some embodiments, the hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) is Molidustat or a pharmaceutically acceptable salt thereof.

[0031] In some embodiments, the hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) is Enarodustat or a pharmaceutically acceptable salt thereof.

[0032] In some embodiments, the hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) is Desidustat or a pharmaceutically acceptable salt thereof.

[0033] In some embodiments, the hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) is HIF117 (SSS17) or a pharmaceutically acceptable salt thereof.

[0034] In some embodiments, the hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) is HEC53856 or a pharmaceutically acceptable salt thereof.

[0035] In some embodiments, the hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) is DDO-3055 or a pharmaceutically acceptable salt thereof.

[0036] In some embodiments, the hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) is a compound of formula I disclosed in WO2018205928 or a pharmaceutically acceptable salt thereof.

[0037] In some embodiments, the hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) is a mixture of any of the above-mentioned HIF-PHIs.

[0038] In this application, unless otherwise indicated, when any compound or pharmaceutically acceptable salt thereof is mentioned or referred to by structural formula, name or code, for example, "compound of Formula I" (synonymous with "compound represented by general formula I"), "rosuvastatin", or "(6'-hydroxy-8'-oxo-3'-phenyl-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carbonyl)glycine", etc., all encompass various forms of these compounds, such as their tautomers, optical isomers, geometric isomers, or isomer mixtures (e.g., racemic mixtures), their isotopically modified compounds, and also encompasses solvates or hydrates of these compounds, as well as their various solid forms (e.g., various crystalline forms or amorphous forms).

[0039] The term "optical isomer" means that when a compound has one or more chiral centers, each chiral center can exist in the R configuration or the S configuration, and the various isomers thus formed are optical isomers. Optical isomers include all diastereomers, enantiomers, meso-isomers, racemates, or mixtures thereof. For example, optical isomers can be separated by chiral chromatography or by chiral synthesis.

[0040] The term "geometric isomer" means that when a compound has a double bond, the compound may exist as cis-isomers, trans-isomers, E-isomers and Z-isomers. Geometric isomers include cis-isomers, trans-isomers, E-isomers, Z-isomers or mixtures thereof.

[0041] The term "tautomer" refers to an isomer resulting from the rapid shift of an atom in a molecule between two positions. Those skilled in the art will appreciate that tautomers can interconvert and, under certain conditions, may reach an equilibrium state and coexist. For example, the term "compound of Formula I" herein also encompasses any tautomer of the compound of Formula I. Specifically, the inventors have discovered that the compound of Formula I may exist as the following tautomers: Ia, Ib, Ic, or Id:

[0042] An "isotopically modified compound" is a compound in which any atom is replaced by an isotope of the compound. The term "isotopically modified compound" as used herein includes all pharmaceutically acceptable isotopically modified compounds of a compound in which one or more atoms are replaced by atoms having the same atomic number as the atoms commonly found in nature but a different atomic mass or mass number.

[0043] Examples of suitable isotopes for inclusion in the compounds of the present application include isotopes of hydrogen such as 2 H(D) and 3 H(T), isotopes of carbon, such as 11 C. 13 C and 14 C, isotopes of chlorine, such as 36 Cl, isotopes of fluorine, such as 18 F, isotopes of iodine, such as 123 I and 125 I, isotopes of nitrogen, such as 13 N and 15 N, isotopes of oxygen, such as 15 O. 17 O and 18 O, and isotopes of sulfur such as 35 S.

[0044] Certain isotopically modified compounds (e.g., those containing radioactive isotopes) are useful in drug and / or substrate tissue distribution studies. The isotope deuterium (i.e., 2 H), tritium (i.e. 3 H) and carbon-14 (i.e. 14 C) is particularly useful for this purpose.

[0045] Using deuterium (i.e.2 H) or tritium (i.e. 3 Substitution with heavier isotopes of H) may afford certain therapeutic advantages resulting from greater metabolic stability (for example, increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances.

[0046] Using positron-emitting isotopes such as 11 C. 18 F. 15 O and 13 N) substitution can be used in positron emission topography (PET) studies to detect substrate receptor occupancy status.

[0047] Isotopically modified compounds can generally be prepared by conventional techniques known to those skilled in the art or by using an appropriate isotopically labeled reagent in place of the non-labeled reagent previously used in analogy to the procedures described in the Examples and Preparations appended herein.

[0048] Certain compounds of the present application can exist in unsolvated forms as well as solvated forms, including hydrated forms.

[0049] Certain compounds of the present application may exist in different crystalline or amorphous forms, and any form thereof is included within the scope of the present application.

[0050] In this application, "pharmaceutically acceptable salts" refer to inorganic or organic acid addition salts or organic or inorganic base addition salts of the compound that are suitable for use in mammals (i.e., with safety and effectiveness when used). These salts can be prepared in situ during the final separation and purification of the compound, or by reacting the pure compound in free form with a suitable organic or inorganic acid or base, and separating the resulting salt. Typical salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, glucoheptonate, lactobionate, laurylsulfonate, etc. These salts can include salts based on alkali and alkaline earth metal cations (e.g., sodium, lithium, potassium, calcium, magnesium, etc.), as well as non-toxic ammonium, quaternary ammonium, and amine cations (including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like).

[0051] In some preferred embodiments of the present application, the hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) is a compound of formula I disclosed in WO2018205928 or a pharmaceutically acceptable salt thereof:

[0052] In some embodiments, in the compound represented by formula I:

[0053] R 1 and R 2 Independently selected from cyano, alkyl, heterocyclic, alkenyl, alkynyl, aryl, heteroaryl and acyl; wherein the above alkyl, heterocyclic, alkenyl, alkynyl, aryl, heteroaryl, acyl are optionally substituted by one or more substituents, wherein the substituents are independently selected from halogen, cyano, hydroxyl, amino, carboxyl, acyl, alkyl, heterocyclic, alkenyl, alkynyl, aryl, heteroaryl, =O, =S, -SH, R 10 O-、R 10 S-、R 10 (O=)S- and R 10 (O=)2S-, where R 10 is alkyl, heterocyclyl, alkenyl, alkynyl, aryl, or heteroaryl; or R 1 and R 2 Put together to form a ring;

[0054] R 3 is selected from alkyl, heterocyclyl, alkenyl, alkynyl, aryl and heteroaryl;

[0055] R 4 and R 5 is hydrogen;

[0056] R 6 and R 6’ is hydrogen;

[0057] R 7 is hydrogen;

[0058] R 8 is selected from hydrogen and alkyl;

[0059] X is an oxygen atom.

[0060] In some embodiments, in the compound represented by formula I:

[0061] R 1 and R 2independently selected from cyano, C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C6-C14 aryl, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, heterocycloalkyl containing 3 to 8 ring atoms, heterocycloalkenyl containing 3 to 8 ring atoms, heteroaryl containing 5 to 14 ring atoms, C1-C12 alkyl-C(=O)- and C2-C12 alkenyl-C(=O)-; wherein the above-mentioned C1-C12 alkyl, C2-C12 alkenyl, C2 -C12 alkynyl, C6-C14 aryl, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, heterocycloalkyl containing 3 to 8 ring atoms, heterocycloalkenyl containing 3 to 8 ring atoms, heteroaryl containing 5 to 14 ring atoms, C1-C12 alkyl-C(=O)-, C2-C12 alkenyl-C(=O)- are optionally substituted with 1 to 3 substituents, wherein the substituents are independently selected from hydroxy, halogen, cyano, amino, carboxyl, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl-, C2-C6 alkynyl-, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, C6-C14 aryl, heteroaryl containing 5 to 14 ring atoms, C1-C6 alkyl-O-, C3-C8 cycloalkyl-O-, C2-C6 alkenyl-O-, C2-C6 alkynyl-O-, C3-C8 cycloalkenyl-O-, C6-C14 aryl-O-, heteroaryl containing 5 to 14 ring atoms, C1-C6 alkyl-S-, C3-C8 cycloalkyl -S-, C2-C6 alkenyl-S-, C2-C6 alkynyl-S-, C3-C8 cycloalkenyl-S-, C6-C14 aryl-S-, heteroaryl-S- containing 5 to 14 ring atoms, heterocycloalkyl containing 3 to 8 ring atoms, heterocycloalkenyl containing 3 to 8 ring atoms, =O, =S, -SH, -CF3, -CO2, C1-C6 alkyl, C1-C6 alkyl-S-, C1-C6 alkyl(O=)S- and C1-C6 alkyl(O=)2S-; or

[0062] R 1 and R 2 taken together to form an optionally substituted cycloalkane, cycloalkene, heterocycloalkane or heterocycloalkene ring containing 3 to 8 ring atoms;

[0063] R 3C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C6-C14 aryl, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, heterocycloalkyl containing 3 to 8 ring atoms, heterocycloalkenyl containing 3 to 8 ring atoms and heteroaryl containing 5 to 14 ring atoms; wherein the above-mentioned C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C6-C14 aryl, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, heterocycloalkyl containing 3 to 8 ring atoms, heterocycloalkenyl containing 3 to 8 ring atoms and heteroaryl containing 5 to 14 ring atoms are optionally substituted with 1 to 3 substituents, wherein the substituents are independently selected from hydroxy, Halogen, cyano, amino, carboxyl, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl-, C2-C6 alkynyl-, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, C6-C14 aryl, heteroaryl containing 5 to 14 ring atoms, C1-C6 alkyl-O-, C3-C8 cycloalkyl-O-, C2-C6 alkenyl-O-, C2-C6 alkynyl-O-, C3-C8 cycloalkenyl-O-, C6-C14 aryl-O-, heteroaryl containing 5 to 14 ring atoms, C1-C6 C6-C6 alkyl-S-, C3-C8 cycloalkyl-S-, C2-C6 alkenyl-S-, C2-C6 alkynyl-S-, C3-C8 cycloalkenyl-S-, C6-C14 aryl-S-, heteroaryl-S- containing 5 to 14 ring atoms, heterocycloalkyl containing 3 to 8 ring atoms, heterocycloalkenyl containing 3 to 8 ring atoms, =O, =S, -SH, -CF3, -CO2, C1-C6 alkyl, C1-C6 alkyl-S-, C1-C6 alkyl(O=)S- and C1-C6 alkyl(O=)2S-;

[0064] R 4 and R 5 is hydrogen;

[0065] R 6 and R 6’ is hydrogen;

[0066] R 7 is hydrogen;

[0067] R 8is selected from hydrogen, C1-C12 alkyl and C3-C8 cycloalkyl; wherein the C1-C12 alkyl and C3-C8 cycloalkyl are optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxy, halogen, cyano, amino, carboxyl, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl-, C2-C6 alkynyl-, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, C6-C14 aryl, heteroaryl containing 5 to 14 ring atoms, C1-C6 alkyl-O-, C3-C8 cycloalkyl-O-, C2-C6 alkenyl-O-, C2-C6 alkynyl-O-, C3-C8 cycloalkenyl-O-, C6-C14 aryl-O- and heteroaryl containing 5 to 14 ring atoms;

[0068] X is an oxygen atom.

[0069] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 are independently selected from cyano and unsubstituted C1-C6 alkyl; or R 1 and R 2 Taken together they form an optionally substituted cycloalkane ring or heterocycloalkane ring containing 3 to 8 ring atoms.

[0070] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 are independently selected from methyl and ethyl; or R 1 and R 2 Together they form a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, a methylcyclohexane ring, a cycloheptane ring, a tetrahydrofuran ring or a tetrahydropyran ring.

[0071] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 Independently selected from cyano and unsubstituted C1-C6 alkyl.

[0072] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 Independently selected from methyl and ethyl.

[0073] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 Taken together they form an optionally substituted heterocycloalkane ring containing 3 to 8 ring atoms.

[0074] In some preferred embodiments of the present application, in the compound represented by formula I, R 1and R 2 Together they form a tetrahydrofuran ring or a tetrahydropyran ring.

[0075] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 Taken together they form an optionally substituted cycloalkane ring containing 3 to 8 ring atoms.

[0076] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 Together they form a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, a methylcyclohexane ring or a cycloheptane ring.

[0077] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 is selected from C1-C6 alkyl, C6-C14 aryl, C3-C8 cycloalkyl and heteroaryl containing 5 to 14 ring atoms; wherein the above-mentioned C1-C6 alkyl, C6-C14 aryl and C3-C8 cycloalkyl are optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxy, halogen, cyano, amino, carboxyl, C6-C14 aryl and C1-C6 alkyl.

[0078] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 is selected from the group consisting of methyl, ethyl, propyl, butyl, phenyl, benzyl, tolyl, methoxyphenyl, chlorophenyl, fluorophenyl, bromophenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and pyridyl.

[0079] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 Selected from C1-C6 chain alkyl; wherein the above-mentioned C1-C6 chain alkyl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxyl, halogen, cyano, amino, carboxyl, C6-C14 aryl and C1-C6 chain alkyl.

[0080] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 Selected from methyl, ethyl, propyl and butyl.

[0081] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 Selected from C6-C14 aryl; wherein the above C6-C14 aryl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxyl, halogen, cyano, amino, carboxyl, C6-C14 aryl and C1-C6 alkyl.

[0082] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 It is selected from phenyl, benzyl, tolyl, methoxyphenyl, chlorophenyl, fluorophenyl, and bromophenyl, preferably selected from phenyl.

[0083] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 Selected from C3-C8 cycloalkyl; wherein the above-mentioned C3-C8 cycloalkyl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxyl, halogen, cyano, amino, carboxyl, C6-C14 aryl and C1-C6 chain alkyl.

[0084] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 Selected from cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0085] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 is selected from heteroaryl groups containing 5 to 14 ring atoms.

[0086] In some preferred embodiments of the present application, in the compound represented by formula I, R 3 Selected from pyridyl.

[0087] In some preferred embodiments of the present application, in the compound represented by formula I, R 8 is selected from hydrogen and C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxy, halogen, cyano, amino, carboxyl, C1-C6 alkyl and C1-C6 alkyl-O-.

[0088] In some preferred embodiments of the present application, in the compound represented by formula I, R 8 Selected from hydrogen, methyl, ethyl, propyl, butyl and pentyl.

[0089] In some preferred embodiments of the present application, in the compound represented by formula I, R 8 Selected from hydrogen.

[0090] In some preferred embodiments of the present application, in the compound represented by formula I, R 8 Selected from C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxy, halogen, cyano, amino, carboxyl, C1-C6 alkyl and C1-C6 alkyl-O-.

[0091] In some preferred embodiments of the present application, in the compound represented by formula I, R 8 Selected from methyl, ethyl, propyl, butyl and pentyl.

[0092] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 taken together to form an optionally substituted cycloalkane ring containing 3 to 8 ring atoms; R 3 is selected from C6-C14 aryl; wherein the above C6-C14 aryl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxyl, halogen, cyano, amino, carboxyl, C6-C14 aryl and C1-C6 alkyl; and R 8 is selected from hydrogen and C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxy, halogen, cyano, amino, carboxyl, C1-C6 alkyl and C1-C6 alkyl-O-.

[0093] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 are each independently selected from C1-C6 alkyl, wherein the above C1-C6 alkyl is optionally substituted with 1 to 3 C1-C6 alkyl substituents; or R 1 and R 2 Taken together to form a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, a methylcyclohexane ring, a cycloheptane ring or a tetrahydropyran ring; R 3 is selected from C6-C14 aryl; wherein the above C6-C14 aryl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxyl, halogen, cyano, amino, carboxyl, C6-C14 aryl and C1-C6 alkyl; and R 8 is selected from hydrogen and C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxy, halogen, cyano, amino, carboxyl, C1-C6 alkyl and C1-C6 alkyl-O-.

[0094] In some preferred embodiments of the present application, in the compound represented by formula I, R 1 and R 2 Together they form a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, a methylcyclohexane ring, and a cycloheptane ring; R 3is selected from C6-C14 aryl; wherein the above C6-C14 aryl is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxyl, halogen, cyano, amino, carboxyl, C6-C14 aryl and C1-C6 alkyl; and R 8 Selected from hydrogen.

[0095] In some particularly preferred embodiments of the present application, the compound represented by general formula I is a compound (6'-hydroxy-8'-oxo-3'-phenyl-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carbonyl)glycine, wherein R 1 and R 2 Together with the C atoms to which they are attached, they form a cyclopentane ring (i.e., R 1 and R 2 -CH2CH2CH2CH2-), R 3 is phenyl, R 4 and R 5 Hydrogen, R 6 and R 6’ is hydrogen, R 7 is hydrogen, R 8 is a compound of formula I wherein R is hydrogen and X is an oxygen atom.

[0096] In the above different preferred embodiments, the preferred options of each substituent can be combined with each other, and various combinations thereof are within the scope of the present application.

[0097] To avoid ambiguity, the following definitions are given for various terms used in this document. Unless otherwise specified, the meanings of the terms used in this document are as follows.

[0098] The term "hydroxy" refers to -OH.

[0099] The term "halogen" or "halo" refers to -F, -Cl, -Br, or -I.

[0100] The term "amino" refers to -NH2.

[0101] The term "cyano" refers to -CN.

[0102] The term "carboxyl" refers to -C(=O)OH.

[0103] The term "substituted" means that one or more (preferably 1 to 5, more preferably 1 to 3) hydrogen atoms in the group are independently replaced by a corresponding number of substituents.

[0104] The term "independently" means that when there are more than one substituent, these substituents may be the same or different.

[0105] The term "optional" or "optionally" means that the event it describes can occur or not occur. For example, a group "optionally substituted" means that the group can be unsubstituted or substituted.

[0106] The term "heteroatom" as used herein refers to oxygen (O), nitrogen (N), or S (O) m (wherein m can be 0, 1 or 2, i.e. a sulfur atom S, or a sulfoxide group SO, or a sulfonyl group S(O)2).

[0107] The term "alkyl" refers to a saturated hydrocarbon composed solely of carbon and hydrogen, formed when any carbon atom loses a hydrogen atom. "Alkyl" as used herein includes chain alkyl groups, such as straight-chain and branched alkyl groups; it also includes cycloalkyl groups, such as monocyclic, spirocyclic, fused, and bridged cycloalkyl groups. Preferably, "alkyl" as used herein refers to chain alkyl groups. Alkyl groups may be unsubstituted or substituted.

[0108] The “alkyl” described herein includes optionally substituted chain alkyl groups, preferably having 1 to 20 carbon atoms, more preferably 1 to 12 carbon atoms, and most preferably 1 to 6 carbon atoms; for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, chloromethyl, fluoroethyl, trifluoromethyl or 1,1,1-trifluoroethyl, and the like.

[0109] The "alkyl" described herein also includes optionally substituted cycloalkyl (e.g., C3-C20 cycloalkyl or C3-C12 cycloalkyl or C3-C8 cycloalkyl), for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, decahydronaphthyl, norpinyl, adamantyl, fluorocyclopropyl, 2-iodocyclobutyl, 2,3-dimethylcyclopentyl, 2,2-dimethoxycyclohexyl and 3-phenylcyclopentyl, etc.

[0110] "C1-C6 chain alkyl" is also called "lower chain alkyl", which refers to a subset of alkyl groups, which refers to straight-chain alkyl groups and branched-chain alkyl groups having 1 to 6 carbon atoms, including, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, etc.

[0111] The "alkyl" described herein is optionally substituted with one or more substituents, wherein the substituents are independently selected from halogen, cyano, nitro (-NO2), hydroxyl, amino, carboxyl, acyl, alkyl, heterocyclyl, alkenyl, alkynyl, aryl, heteroaryl, =O, =S, -SH, R 16 O-、R 16 S-、R 16 (O=)S-、R 16 (O=)2S-, where R 16is an alkyl group, a heterocyclyl group, an alkenyl group, an alkynyl group, an aryl group, or a heteroaryl group.

[0112] Preferably, the "alkyl" described herein is optionally substituted by 1 to 3 substituents, wherein the substituents are independently selected from hydroxy, halogen, nitro, cyano, amino, carboxyl, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl-, C2-C6 alkynyl-, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, C6-C14 aryl, heteroaryl containing 5 to 14 ring atoms, C1-C6 alkyl-O-, C3-C8 cycloalkyl-O-, C2-C6 alkenyl-O-, C2-C6 alkynyl-O-, C3-C8 cycloalkenyl-O-, C6-C14 aryl- O-, heteroaryl-O- containing 5 to 14 ring atoms, C1-C6 alkanyl-S-, C3-C8 cycloalkyl-S-, C2-C6 alkenyl-S-, C2-C6 alkynyl-S-, C3-C8 cycloalkenyl-S-, C6-C14 aryl-S-, heteroaryl-S- containing 5 to 14 ring atoms, heterocycloalkyl containing 3 to 8 ring atoms, heterocycloalkenyl containing 3 to 8 ring atoms, =O, =S, -SH, -CF3, -CO2C1-C6 alkanyl, C1-C6 alkanyl-S-, C1-C6 alkanyl(O=)S- and C1-C6 alkanyl(O=)2S-

[0113] The term "alkenyl" refers to any hydrocarbon group consisting solely of carbon and hydrogen, containing one or more carbon-carbon double bonds but no carbon-carbon triple bonds or aromatic bonds, formed when any carbon atom loses a hydrogen atom. "Alkenyl" as used herein includes alkenyl groups, such as straight-chain alkenyl and branched-chain alkenyl groups, as well as cycloalkenyl groups, such as monocyclic alkenyl, spirocyclic alkenyl, fused-cycle alkenyl, and bridged-cycle alkenyl groups. Alkenyl groups may be unsubstituted or substituted.

[0114] The “alkenyl” group described herein includes optionally substituted alkenyl groups, preferably having 2 to 20 carbon atoms, more preferably having 2 to 12 carbon atoms, and most preferably having 2 to 6 carbon atoms; for example, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, isobutenyl, 1-pentenyl, 2-pentenyl, isopentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, isohexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 1-octenyl, 2-octenyl, 3-octenyl, 4-octenyl, 1-nonenyl, 1-decenyl, 1-undecenyl, 1-dodecenyl, and the like.

[0115] The term "alkenyl" described herein also includes optionally substituted cycloalkenyl groups (e.g., C3-C20 cycloalkenyl or C3-C12 cycloalkenyl or C3-C8 cycloalkenyl), for example, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclobutadienyl, cyclopentadienyl, cycloheptatrienyl, etc.

[0116] The "alkenyl" described herein is optionally substituted with one or more (eg, 1-3) substituents, wherein the selection and preferred range of the substituents are the same as those of the "alkyl" substituents.

[0117] The term "alkynyl" refers to a group formed when any carbon atom of a hydrocarbon composed solely of the elements carbon and hydrogen, containing one or more carbon-carbon triple bonds but no aromatic bonds, loses a hydrogen atom. "Alkynyl" as used herein includes chain alkynyl groups, such as straight-chain and branched alkynyl groups, as well as cycloalkynyl groups, such as monocyclic, spirocyclic, fused, and bridged cyclic alkynyl groups. Alkynyl groups may optionally contain one or more carbon-carbon double bonds. Alkynyl groups may be unsubstituted or substituted.

[0118] The “alkynyl” described herein includes optionally substituted alkynyl groups, preferably having 2 to 20 carbon atoms, more preferably having 2 to 12 carbon atoms, and most preferably having 2 to 6 carbon atoms; for example, ethynyl, propynyl, 1-butynyl, 2-butynyl, isobutynyl, 1-pentynyl, 2-pentynyl, isopentenyl, 3-methyl-3-butynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 3-heptynyl, 1-octynyl, 1-nonynyl, 1-decynyl, 1-undecanynyl, 1-dodecynyl, and the like.

[0119] The term "alkynyl" described herein also includes optionally substituted cycloalkynyl (eg, C8-C18 cycloalkynyl), for example, cyclooctynyl and the like.

[0120] The "alkynyl" described herein is optionally substituted by one or more (eg, 1-3) substituents, wherein the selection and preferred range of the substituents are the same as those of the "alkyl" substituents.

[0121] The term "heterocyclyl" refers to a substituent of a monocyclic or polycyclic compound that is saturated or contains a carbon-carbon double bond or a carbon-carbon triple bond; it contains 3 to 20 ring atoms (preferably 3 to 12 ring atoms, more preferably 3 to 8 ring atoms), one or more of which are selected from heteroatoms and the remaining ring atoms are carbon; and none of the rings are aromatic. The "heterocyclyl" described herein also includes spiroheterocyclyl, fused heterocyclyl, and bridged heterocyclyl. The heterocyclyl group may be unsubstituted or substituted. The heterocyclyl group may be a heterocycloalkyl, heterocycloalkenyl, or heterocycloalkynyl. Examples of suitable monocyclic heterocycles include, but are not limited to, piperidinyl, pyrrolidinyl, piperazinyl, azetidinyl, aziridine, morpholinyl, thietanyl, oxolanyl (tetrahydrofuranyl), oxahenyl (tetrahydropyranyl), and the like.

[0122] It should be understood that the "heterocyclyl" described herein is optionally substituted by one or more (eg, 1-3) substituents, wherein the selection and preferred range of the substituents are the same as those of the "alkyl" substituents.

[0123] The term "aryl" refers to a 6- to 14-membered, all-carbon monocyclic or fused polycyclic ring having a conjugated π-electron system. The aryl ring may be fused with a heteroaromatic ring, a heterocyclic ring, a cycloalkane, a spirocycloalkane, a fused cycloalkane, a bridged cycloalkane, a cycloalkene, a spirocycloalkene, a fused cycloalkene, a bridged cycloalkene, a cycloalkyne, a spirocycloalkyne, a fused cycloalkyne, or a bridged cycloalkyne. The aryl group may be unsubstituted or substituted. Examples include, but are not limited to, phenyl, naphthyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, methoxyphenyl (e.g., 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl), chlorophenyl (e.g., 2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl), fluorophenyl (e.g., 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl), bromophenyl (e.g., 2-bromophenyl, 3-bromophenyl, 4-bromophenyl), 2-chloro-3-methylphenyl, 2-chloro-4-methylphenyl, 2-chloro-5-methylphenyl, 3-chloro-2-methylphenyl, 3-chloro-4-methylphenyl, 4-chloro-2-methylphenyl, 4-chloro-3-methylphenyl, 5-chloro-2-methylphenyl, 2,3-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 2,3-dimethylphenyl, 3,4-dimethylphenyl, and the like.

[0124] The "aryl" described herein is optionally substituted by 1-4 or 1-3 substituents, wherein the selection and preferred range of the substituents are the same as those of the "alkyl" substituents.

[0125] The term "heteroaryl" refers to an aromatic system comprising 5 to 18 ring atoms (preferably 5 to 14 ring atoms) wherein 1 to 4 of the ring atoms are heteroatoms selected from oxygen, nitrogen, and sulfur. The heteroaryl group itself may be fused to an aromatic ring, a heterocycle, a cycloalkane, a spirocycloalkane, a fused cycloalkane, a bridged cycloalkane, a cycloalkene, a spirocycloalkene, a fused cycloalkene, a bridged cycloalkene, a cycloalkyne, a spirocycloalkyne, a fused cycloalkyne, or a bridged cycloalkyne. It may be unsubstituted or substituted. Examples of heteroaryl groups include, but are not limited to, thienyl, furyl, pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, pyrazinyl, oxazolyl, thiazolyl, benzothienyl, benzofuranyl, benzoxazolyl, benzimidazolyl, indolyl, quinolyl, isoquinolyl, and quinazolinyl.

[0126] The "heteroaryl" described herein is optionally substituted by 1-4 or 1-3 substituents, wherein the selection and preferred range of the substituents are the same as those of the "alkyl" substituents.

[0127] As used herein, the term "acyl" refers to RC(=O)-, wherein R is C1-C18 (preferably C1-C12, more preferably C1-C6) alkyl. Examples of "acyl" include, but are not limited to, formyl, acetyl, benzoyl, nicotinoyl, propionyl, isobutyryl, oxalyl, and the like.

[0128] The acyl group RC(=O)- is optionally substituted by one or more (eg, 1-3) substituents, wherein the selection and preferred range of the substituents are the same as those of the "alkyl" group.

[0129] The term "cyclization" as used herein refers to the formation of a cyclic structure such as a cycloalkane ring, a cycloalkene ring, a cycloalkyne ring, an aromatic ring, a heterocycloalkane ring, a heterocycloalkene ring, a heterocycloalkyne ring, a heteroaromatic ring, etc. The cyclic structure may be a monocyclic, bicyclic or polycyclic structure, including fused rings, bridged rings, and spirocyclic structures thereof. In particular, the substituent R herein 1 and R 2 The formed ring is preferably a 3- to 12-membered ring, particularly preferably a 3- to 12-membered cycloalkane ring, cycloalkene ring, heterocycloalkane ring, heterocycloalkene ring, and most preferably a 3- to 8-membered cycloalkane ring, cycloalkene ring, heterocycloalkane ring, heterocycloalkene ring, such as a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, a cycloheptane ring, a tetrahydrofuran ring, a tetrahydropyran ring, etc. The ring structure is optionally substituted with one or more (e.g., 1-3) substituents, wherein the selection and preferred range of the substituents are the same as those for the substituents of the "alkyl" group.

[0130] In this article, the numerical ranges related to the number of substituents, the number of carbon atoms, and the number of ring atoms represent a list of all integers within the range, and the range is only used as a simplified representation. For example:

[0131] "1-4 substituents" means 1, 2, 3 or 4 substituents;

[0132] "1-3 substituents" means 1, 2 or 3 substituents;

[0133] "3- to 12-membered ring" means a 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered ring;

[0134] "3- to 8-membered ring" means a 3-, 4-, 5-, 6-, 7- or 8-membered ring;

[0135] “1-12 carbon atoms” or “C1-C12” means 1 (C1), 2 (C2), 3 (C3), 4 (C4), 5 (C5), 6 (C6), 7 (C7), 8 (C8), 9 (C9), 10 (C10), 11 (C11) or 12 carbon atoms (C12);

[0136] “1-6 carbon atoms” or “C1-C6” means 1 (C1), 2 (C2), 3 (C3), 4 (C4), 5 (C5) or 6 carbon atoms (C6);

[0137] “2-6 carbon atoms” or “C2-C6” means 2 (C2), 3 (C3), 4 (C4), 5 (C5) or 6 carbon atoms (C6);

[0138] "C3-C8" means 3 (C3), 4 (C4), 5 (C5), 6 (C6), 7 (C7) or 8 carbon atoms (C8);

[0139] "3 to 8 ring atoms" means 3, 4, 5, 6, 7 or 8 ring atoms.

[0140] Therefore, numerical ranges related to the number of substituents, the number of carbon atoms, and the number of ring atoms also encompass any subranges thereof, and each subrange is also considered to be disclosed herein.

[0141] In some particularly preferred embodiments of the present application, the hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) is a compound disclosed in WO2018205928 (6'-hydroxy-8'-oxo-3'-phenyl-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carbonyl)glycine or a pharmaceutically acceptable salt thereof. The compound (6'-hydroxy-8'-oxo-3'-phenyl-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carbonyl)glycine is wherein R 1 and R 2 Together they form a cyclopentane ring (ie -CH2CH2CH2CH2-), R 3 is phenyl, R 4 and R 5 Hydrogen, R 6 and R 6’ is hydrogen, R 7 is hydrogen, R 8 is a compound of formula I wherein R is hydrogen and X is an oxygen atom.

[0142] 2. Drug Use or Treatment Methods

[0143] A first aspect of the present application provides use of a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) in the manufacture of a medicament for treating rare anemias.

[0144] A second aspect of the present application provides the use of a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) in the manufacture of a medicament for increasing red blood cell count, hemoglobin content and / or hematocrit in rare anemias.

[0145] The third aspect of the present application provides a method for treating rare anemias, comprising: administering a therapeutically effective amount of a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) to a patient in need of treatment.

[0146] The fourth aspect of the present application provides a method for increasing the red blood cell count, hemoglobin content and / or hematocrit of rare anemias, comprising: administering a therapeutically effective amount of a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) to a patient in need of treatment.

[0147] The fifth aspect of the present application provides a hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) for treating rare anemias.

[0148] The sixth aspect of the present application provides the use of hypoxia-inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) for treating rare anemias.

[0149] As used herein, the term "patient" refers to all mammals, including humans. Examples of patients include humans, cows, dogs, cats, goats, sheep, mice, pigs, rabbits, rats, or mice.

[0150] Herein, the rare anemia may be selected from myelodysplastic syndrome anemia (MDS anemia), beta-thalassemia (β-thalassemia) or sickle cell disease (sickle cell anemia or SCD anemia).

[0151] The hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) of the present application can also be used to treat anemia complications or ischemic diseases, such as ischemic cerebrovascular disease (ischemic cerebrovascular disease), ischemic renal disease (IRD), ischemic cardiomyopathy (ICM), etc.

[0152] When using the hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) of the present application to treat rare anemias, it is generally necessary to administer a therapeutically effective amount of the active ingredient (i.e., HIF-PHI) to the patient. As used herein, the term "therapeutically effective amount" refers to an amount that, when the active ingredient mentioned in the present application is administered to the patient, can effectively delay or eliminate the patient's symptoms or improve the patient's health. The specific dosage used can be determined by the doctor based on the patient's specific circumstances. The precise dosage to be used depends not only on the route of administration, the disease, the severity of the disease to be treated, and various physical factors associated with the individual being treated, but can also be determined based on the judgment of the health care practitioner. In vitro or in vivo tests can be used to help determine the optimal dosage range.

[0153] For example, the compound of formula I of the present application can be administered to a patient at a dosage of about 0.01-4000 mg, or 0.05-2000 mg, or 0.1-1000 mg, or 0.1-500 mg, or 5-500 mg, 5-300 mg, 5-200 mg, 5-100 mg, 10-30 mg, 5-15 mg, 5-10 mg, 10-15 mg, or 30-50 mg per day. However, the specific dosage used can vary. Those skilled in the art will know how to determine the optimal dosage for a particular patient.

[0154] For therapeutic use, HIF-PHI is usually administered to a patient in the form of a pharmaceutical composition comprising at least one of the above compounds as an active ingredient, optionally together with pharmaceutically acceptable adjuvants and / or excipients and pharmaceutically acceptable solid or liquid carriers.

[0155] The pharmaceutical composition of the present application can be formulated into various pharmaceutical dosage forms applicable to oral administration as required. Therefore, if solid carriers are used, the preparation can be in tablets, placed in hard gel capsules in powder or granular form, or in the form of lozenges or lozenges. Solid carriers can include conventional excipients, such as adhesives, fillers, tableting lubricants, disintegrants, wetting agents, etc. If necessary, the tablets can be coated with conventional technology films. If liquid carriers are used, the preparation can be in the form of syrup, emulsion, soft gel capsules, aqueous or non-aqueous liquid suspensions, or can be the dry product reconstituted with water or other suitable carriers before use. Liquid preparations can include conventional additives, such as suspending agents, emulsifiers, wetting agents, non-aqueous carriers (including edible oils), preservatives, and flavoring agents and / or coloring agents.

[0156] These pharmaceutical compositions (or pharmaceutical preparations) can also include various excipients, for example, preservatives, wetting agents, emulsifiers and dispersants. The inhibition of the effect of microorganisms can be ensured by various antibacterial and antifungal agents (for example, parabens, chlorobutanol, phenol, sorbic acid, etc.). Isotonic agents can also be included, for example, sugar, sodium chloride, etc. The absorption of injectable pharmaceutical dosage forms can be extended by using delayed absorption agents (for example, aluminum monostearate and gel).

[0157] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one inert excipient (or carrier) (e.g., sodium citrate or dicalcium phosphate) and may also include: (a) fillers or admixtures (e.g., starch, lactose, sucrose, glucose, mannitol and silicic acid); (b) binders (e.g., carboxymethylcellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic); (c) humectants (e.g., glycerol); (d) disintegrants (e.g., agar); (e.g., fat-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain synthetic silicates, sodium carbonate); (e) solution retarding agents (e.g., paraffin); (f) absorption promoters (e.g., quaternary ammonium compounds); (g) wetting agents (e.g., cetyl alcohol and glycerol monostearate); (h) adsorbents (e.g., kaolin and bentonite) and (i) lubricants (e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate), or mixtures thereof.

[0158] Solid pharmaceutical compositions (or pharmaceutical formulations) of a similar type may also be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose and high molecular weight polyethylene glycols.

[0159] Solid dosage forms (e.g., tablets, dragees, capsules, pills, and granules) can be prepared with coatings and shells (e.g., enteric coatings and others known in the art). They may contain opacifiers, and they may also be pharmaceutical compositions (or pharmaceutical preparations) that release the active compound or compounds in a delayed manner in a certain part of the intestinal tract. The active ingredient may also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0160] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, dispersions, syrups, and elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents (e.g., water or other solvents), solubilizers and emulsifiers (e.g., ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide), oils (specifically, cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil), glycerol, tetrahydrofuran alcohol, polyethylene glycol, and fatty acid esters of sorbitan, or mixtures thereof, etc., commonly used in the art.

[0161] In addition to these inert diluents, the pharmaceutical composition (or pharmaceutical preparation) may also include, for example, wetting agents, emulsifying and suspending agents, perfuming agents, flavoring agents, and perfuming agents.

[0162] In addition to the active compounds, suspensions may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances.

[0163] The amount of the HIF-PHI compound in the pharmaceutical composition (or pharmaceutical preparation) can be appropriately determined by those skilled in the art according to needs.

[0164] In some embodiments, the amount of HIF-PHI or a pharmaceutically acceptable salt thereof contained in each unit of the pharmaceutical preparation is: about 1-1000 mg, about 1-10 mg, about 10 mg, about 5 mg, about 4.5 mg, about 1-3 mg, about 2-4 mg, about 3-5 mg, about 4-6 mg, about 5-7 mg, about 6-8 mg, about 7-9 mg, about 8-10 mg, about 9-11 mg, about 10-12 mg, about 4.5-5 mg, 20 mg, 30 mg, 30 -100 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 10-30 mg, about 30-50 mg, about 50-70 mg, about 10-90 mg, about 10-800 mg, about 10-700 mg, about 10-600 mg, about 10-500 mg, about 10-400 mg, about 10-300 mg, or about 10-200 mg, or an amount within any range defined by any of these values.

[0165] In some embodiments, the amount of HIF-PHI or a pharmaceutically acceptable salt thereof contained in each unit of the pharmaceutical preparation is about 0.01-0.2 mg, about 0.2-0.4 mg, about 0.4-0.6 mg, about 0.6-0.8 mg, about 0.8-1 mg, about 1-1.2 mg, about 1.2-1.4 mg, about 1.4-1.6 mg, about 1.6-1.8 mg, about 1.8-2 mg, about 2-2.2 mg, about 2.2-2.4 mg, about 2.4-2.6 mg, about 2.6-2.8 mg, about 2.8-3 mg, about 3-3.2 mg, about 3.2-3.4 mg, about 3.4-3.6 mg, about 3.6-3.8 mg, about 3.8-4 mg, about 3.4-3.6 mg. 9-4.1mg, about 4-4.2mg, about 0.2-0.4mg, about 0.2-0.6mg, about 0.2-0.8mg, about 0.2-1mg, about 0.2-1.2mg, about 0.2-1.4mg, about 0.2-1.6mg, about 0.2-1.8mg, about 0.2-2.0mg, 0.2-2.5mg, about 0.2-3.0mg, about 0.2-3.5mg, about 0.2-4.0mg, about 5-10mg, about 10-15mg, about 15-20mg, about 20-25mg, about 25-30mg, about 30-40mg, about 40-50mg, about 50-60mg, about 60-70mg, about 70-80mg, about 80- 90mg, about 90-100mg, about 100-120mg, about 120-140mg, about 140-150mg, about 150-160mg, about 160-180mg, about 180-200mg, about 200-220mg, about 220-240, about 10-500mg, about 50-400mg, about 50-300mg, about 100-250mg, about 1-10mg, about 10-200mg, about 10-150mg, about 10-100mg, about 10-180mg, about 10-160mg, about 10-140mg, about 10-120mg, about 10-100mg, about 10-20mg, about 20-30mg, about 3 In some embodiments, the present invention relates to an amount of at least one pharmaceutical composition comprising: a ...

[0166] In some embodiments, the amount of HIF-PHI or a pharmaceutically acceptable salt thereof contained in each unit of the pharmaceutical preparation is in the following proportions based on the total weight of the pharmaceutical preparation: about 0.1% (w / w) to about 10% (w / w), about 0.05% (w / w) to about 5% (w / w), about 0.2% (w / w) to about 15% (w / w), about 0.5% (w / w) to about 30% (w / w), about 1% (w / w) to about 25% (w / w), about 2% (w / w), about 3% (w / w), about 4% (w / w), about 5% (w / w), at least about 10% (w / w), at least about 20% (w / w), at least about 50% (w / w), at least about 70% (w / w). % (w / w), at least about 80%, about 10% (w / w) to about 30% (w / w), about 10% (w / w) to about 20% (w / w), about 20% (w / w) to about 30% (w / w), about 30% (w / w) to about 50% (w / w), about 30% (w / w) to about 40% (w / w), about 40% (w / w) to about 50% (w / w), about 50% (w / w) to about 80% (w / w), about 50% (w / w) to about 60% (w / w), about 70% (w / w) to about 80% (w / w), or about 80% (w / w) to about 90% (w / w), or a ratio within any range defined by any of these ratios.

[0167] Pharmaceutical compositions (or pharmaceutical preparations) can be prepared by methods known or commonly used in the art, which generally include mixing the active ingredient HIF-PHI with a pharmaceutically acceptable carrier, adjuvant, or excipient, and optional post-treatment or processing steps (e.g., drying, granulation, encapsulation, etc.).

[0168] Example

[0169] The embodiments described below are some embodiments of the present invention, rather than all embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without departing from the principles of the present invention and without making creative efforts are within the scope of protection of the present invention.

[0170] For the sake of clarity, some materials, equipment, and methods commonly used in the art are not listed in the examples. Any process methods and analytical testing procedures (and related parameters) not specifically noted in the examples were performed according to those commonly employed by those skilled in the art. Materials, reagents, and equipment not specifically sourced are all routine laboratory materials and equipment and are commercially available.

[0171] Example 1

[0172] Synthesis of (6'-hydroxy-8'-oxo-3'-phenyl-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carbonyl)glycine (1)

[0173] Compound 1 was synthesized according to the method of Example 24 in WO2018205928.

[0174] The structures of the compounds were determined by liquid chromatography-mass spectrometry (LCMS) or nuclear magnetic resonance (NMR). NMR chemical shifts (δ) were expressed in 10 -6 The unit of concentration is expressed in parts per million (ppm). NMR measurements were performed using a Bruker-500 NMR spectrometer, using deuterated dimethyl sulfoxide (DMSO-d6) or deuterated chloroform (CDCl3) as solvents, and tetramethylsilane (TMS) as the internal standard. LCMS measurements were performed using a Shimadzu LCMS-2020 or Thermo UltiMate 3000.

[0175] Thin layer chromatography silica gel plates used were HSGF254 from Yantai Huanghai, Shandong Province or GF254 from Qingdao, Shandong Province. Column chromatography generally used 200 to 300 mesh silica gel from Yantai Huanghai, Shandong Province as a carrier.

[0176] All starting materials used in this example were purchased from chemical suppliers or synthesized according to literature methods.

[0177] The following abbreviations may be used in this example: DMSO-d6: dimethyl sulfoxide with all six hydrogen atoms replaced by deuterium CDCl3: deuterated chloroform CAS: Chemical Abstracts Service NMR: nuclear magnetic resonance LCMS: liquid chromatography-mass spectrometry ESI: electrospray ionization ppm: parts per million δ: nuclear magnetic resonance chemical shift TMS: tetramethylsilane s: nuclear magnetic resonance singlet d: nuclear magnetic resonance doublet t: nuclear magnetic resonance triplet br: nuclear magnetic resonance broad peak CDI: carbonyldiimidazole DCC: N,N'-dicyclohexylcarbodiimide NBS: N-bromocyclosuccinimide

[0178] The main synthesis steps of compound 1 are as follows.

[0179] first step

[0180] 1-(2-Phenyl-1H-pyrrol-1-yl)cyclopentane-1-carboxylic acid (1a)

[0181] Compound 3-(1,3-dioxan-2-yl)-1-phenylpropan-1-one (prepared according to the method described in patent document WO / 2011 / 042477) (500 mg) and 1-amino-cyclopentane-1-carboxylic acid (421 mg) were refluxed in 8 ml of acetic acid for 16 hours. After the reaction was cooled, the acetic acid was evaporated to a minimum, diluted with water and ethyl acetate, and the organic phase was collected and washed several times with dilute sodium chloride solution until the aqueous phase was nearly neutral. The ethyl acetate layer was then dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 1a. LCMS ESI(+): 256 (M+1). + . 1 H NMR(500MHz,DMSO-d6)δ(ppm):12.95(br,s,1H),7.35-7.33(m,3H),7.25-7.22(m,2H),7.00-6 .99(m,1H),6.04(m,1H),5.95(m,1H),2.14-2.11(m,2H),2.01-1.97(m,2H),1.60-1.57(m,4H).

[0182] Step 2

[0183] Dimethyl 2-(1-(2-phenyl-1H-pyrrol-1-yl)cyclopentane-1-carbonyl)malonate (1b)

[0184] The synthetic route of compound 15b in the second step of Example 15 in WO2018205928 was adopted, and the raw material 15a was replaced with 1a to obtain 1b. LCMS ESI (+): 370 (M+1) + . 1 H NMR (500MHz, DMSO-d6) δ7.28–7.22(m,3H),7.20–7.15(m,2H),7.11(dd,J=3.2,1.8Hz,1H),6.15(t,J=3.3Hz,1H),5.93(d d,J=3.5,1.7Hz,1H),5.13(s,1H),3.56(s,6H),2.06(d,J=8.0Hz,2H),1.59(t,J=12.2Hz,2H),1.33(s,2H),1.16(s,2H).

[0185] Step 3

[0186] 6'-Hydroxy-8'-oxo-3'-phenyl-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carboxylic acid methyl ester (1c)

[0187] The synthesis route of compound 15c in the third step of Example 15 in WO2018205928 was adopted, and the raw material 15b was replaced with 1b to obtain 1c. LCMS ESI (+): 338 (M+1) + . 1 H NMR(500MHz,DMSO-d6)δ13.84(s,1H),7.54-7.43(m,5H),7.09(d,J=4.0Hz,1H),6.35(d,J=4.0H z,1H),3.81(s,3H),2.30-2.21(m,2H),2.10-2.01(m,2H),1.48-1.38(m,2H),1.00-0.89(m,2H).

[0188] Step 4

[0189] (6'-Hydroxy-8'-oxo-3'-phenyl-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carbonyl)glycine (1)

[0190] Compound 1 was prepared by using the synthetic route of compound 15 in the fourth step of Example 15 in WO2018205928, replacing the raw material 15c with 1c. LCMS ESI (+): 381 (M+1) + . 1 H NMR (500MHz, DMSO-d6) δ (ppm) 17.96 (s, 1H), 12.97 (s, 1H), 9.89-9.87 (d, J = 5.5Hz, 1H), 7.53-7.46 (m, 5H), 7.08-7.07 (d, 1H) ,6.37-6.36(d,1H),4.08-4.07(d,J=5.5Hz,2H),2.35-2.20(m,2H),2.18-2.10(m,2H),1.60-1.46(m,2H),0.98-0.85(m,2H).

[0191] Example 2

[0192] Inhibitory effects of compounds on HIF PHD-2

[0193] In this example and the following examples, Compound 1 prepared in Example 1 was used for testing, and “Comp-1” was used to refer to Compound 1.

[0194] The following stock solutions were prepared: HEPES buffer (pH 7.4, 50 mM), DMSO / HEPES solution of HIF-1α peptide (sequence: DLDLEMLAPYIPMDDDFQL; Genscript) (0.5 mM; preparation method: first prepare a 20 mM DMSO solution of the peptide, then dilute to 40 times the volume with the aforementioned HEPES buffer), ascorbic acid solution (10 mM), FeSO4 hydrochloric acid solution (0.1 mM, hydrochloric acid concentration 10 μM), 2-oxoglutaric acid solution (2-OG: 1 mM), EDTA solution (pH 7.4, 50 mM), and 1 mM EDTA solution. 8.0; 0.5M), and eight concentrations of Comp-1 / DMSO solution (4000μM, 1333μM, 444.4μM, 148.1μM, 49.4μM, 16.5μM, 5.5μM, and 1.82μM, respectively). Nine Eppendorf tubes were each added with the following reagents: 50μL HEPES solution, 10μL ascorbic acid solution, 10μL FeSO4 hydrochloric acid solution, 10μL 2-OG solution, and 10μL HIF-1α peptide solution. 5μL of each of the eight concentrations of Comp-1 / DMSO solution and 5μL DMSO were then added. Finally, 5μL of PHD2 enzyme (Active Motif; concentration: 0.2μg / μL) was added to each tube. The reaction system was incubated in the dark at 30°C for 2 hours. Finally, 10μL of EDTA solution was added to each tube to terminate the reaction. The liquid of each reaction system was filtered with a 0.45 μm filter, and the HIF-peptide and hydroxylated HIF-peptide were separated by high performance liquid chromatography. The concentrations of HIF-peptide and hydroxylated HIF-peptide were quantified using a UV absorption detector, and their ratio was calculated. IC was calculated using Graph Prism 6.0 software. 50 ; IC of Comp-1 to HIF PHD-2 50 It is 2.36μM.

[0195] Example 3

[0196] Effects of HIF-PH inhibitor and EPO on MDS model mice

[0197] Throughout the experimental period, animals were housed in cages with 24-hour central air conditioning, a temperature set at 20-26°C, a relative humidity of 40-70%, and a 12-hour light-dark cycle. All animals had free access to food and sterile drinking water. The C57 mice used in the experiments contained three knock-in genes: Vav1-Cre, LSL, and c-Myc (T58A / S62E). Animals were required to be positive for all three genes. Vav1-Cre expression induces cleavage of the LSL locus, leading to tTA gene expression. Following induction with doxycycline, tTA binds to the c-Myc (T58A / S62E) promoter, driving c-Myc (T58A / S62E) expression and resulting in the development of the MDS phenotype. Mice were established at approximately 8 weeks of age and fed a specialized diet (200 mg of doxycycline per kg of diet) for 4 weeks. On days 0, 14, and 28, mice were anesthetized and 70 μl of blood was collected from the eyes. Blood levels of red blood cells (RBCs), hemoglobin, and hematocrit (HCT) were measured to confirm successful MDS model establishment. Mice were then randomly divided into three groups of 10 mice each: a vehicle control group (0.1 M meglumine), a low-dose Comp-1 group (10 mg / kg), and a high-dose Comp-1 group (30 mg / kg). The volume of drug administration or vehicle was 6.67 mL / kg. All groups received oral gavage three times per week (TIW) for 4 consecutive weeks, with weeks 4 to 8 representing the treatment period. On days 14 (week 6) and 28 (week 8) of treatment, mice were anesthetized and 70 μl of blood was collected from the eyes. Blood samples were collected in heparinized tubes and assayed for red blood cell (RBC), hemoglobin (Hb), and hematocrit (HCT) content. After the final blood draw, each animal was euthanized and the corresponding organs and tissues were collected and stored for future use. The instrument used was an Element HT5 Veterinary Hematology Analyzer (supplier: HESKA). Graphpad Prism 9.0 was used for data analysis.

[0198] Experimental results:

[0199] (1) Effect on RBC: After oral administration of Comp-1, the RBC in the blood increased significantly. After 2 weeks of administration, the RBC values ​​of 30 mg / kg and 10 mg / kg were significantly higher than those of the vehicle group (7.11M / μL vs 6.33M / μL vs 5.57M / μL). After 4 weeks of administration, the RBC in the treatment group continued to increase (8.46M / μL vs 7.17M / μL vs 5.27M / μL), and the RBC of the 30 mg / kg group animals returned to a level close to that before modeling (8.46M / μL vs 9.01M / μL). See the table below:

[0200] (2) Effect on Hb: After oral administration of Comp-1, the Hb of MDS mice increased significantly. After 2 weeks of administration, the Hb values ​​of 30 mg / kg and 10 mg / kg were significantly higher than those of the vehicle group (13.56 g / dL vs 10.97 g / dL vs 8.72 g / dL). After 4 weeks of administration, the Hb of the treatment group continued to increase (14.91 g / dL vs 11.77 g / dL vs 8.19 g / dL), and the Hb of the 30 mg / kg group returned to the level before modeling (14.91 g / dL vs 14.37 g / dL). See the table below:

[0201] (3) Effect on HCT hematocrit: After oral administration of Comp-1, the HCT of MDS mice increased significantly. After 2 weeks of administration, the HCT values ​​of 30 mg / kg and 10 mg / kg were significantly higher than those of the vehicle group (43.13% vs 34.36% vs 27.69%). After 4 weeks of administration, the HCT of the treatment group continued to rise (47.41% vs 37.52% vs 27.13%), and the HCT of the 30 mg / kg group was even slightly higher than the pre-modeling level after recovery (47.41% vs 43.32%). See the table below:

[0202] It can be seen that the test substance Comp-1 at 10 and 30 mg / kg can effectively increase the RBC, Hb, and HCT values ​​in the blood of MDS mice and significantly improve the anemia symptoms of MDS mice.

[0203] As a control, the same experiment was repeated using Roxadustat (30 mpk, oral gavage, TIW / three times a week) and EPO (50 U / 30 g). Neither Roxadustat nor EPO could improve the RBC, Hb, and HCT values ​​of this MDS mouse model.

[0204] Example 4

[0205] Effects of Comp-1 on homozygous β-thalassemia mice

[0206] During the entire experimental period, the animals were housed in an animal room with 24-hour continuous central air conditioning, a temperature set at 20-26 degrees Celsius, an air relative humidity set at 40-70%, and a 12-hour light-dark cycle. All animals had free access to food (irradiated sterilized feed) and water (prepared by an ultrapure water dispenser). The mouse strain used in the experiment was: Hbbd 3th / d3thMice; male and female, 9-15 weeks old, weighing 25-30 grams. One male is mated with two females for breeding, and the male genotype is Hbb. d3th / d3th The female mice were selected with the genotype Hbb d3th / + Mice. When the offspring mice were one week old, the toes were clipped to extract tissue DNA, which was subsequently amplified by PCR and separated by agarose gel electrophoresis for the purpose of genetic identification. Before the experiment, a secondary genotyping of mice of appropriate age was performed, and mice with no significant differences in weight and appearance were selected as experimental mice. Solvent 1 (0.1M meglumine aqueous solution): An appropriate amount of meglumine (0.586g) was dissolved in an appropriate amount of deionized water (30mL) and stirred to prepare the solvent. After the solvent was prepared, it was divided into daily doses and stored at 5±3°C. Stir at room temperature for at least 10 minutes before administration and continue stirring during administration.

[0207] Comp-1: Weigh the required amount of Comp-1, dissolve it completely in an appropriate volume of solvent 1, and stir and sonicate to form a solution. After dissolution is observed with the naked eye, gradually add the solvent until the specified volume is reached. Before daily administration, take the preparation (including the solvent) out of the refrigerator and allow it to reach room temperature. Stir the preparation at room temperature for at least 10 minutes before administration and stir it throughout the administration process. Information such as animal grouping, dosage, and experimental time length is shown in the table below. The route of administration is oral gavage, the dosage volume is 10 mL / kg, and the administration frequency is daily for a total of 42 days.

[0208] CBC test

[0209] On days 0, 15, 29, and 43 of dosing, 50 μl of blood was collected from the mice via the orbital orbital approach, stored at 4°C, and brought to room temperature before testing. The blood was then gently mixed and added to the prepared diluent (blood:diluent = 1:6). Data were analyzed using the instrument's pre-dilution mode. Data were processed and analyzed using Microsoft Office Excel (Microsoft Corporation, version 2013, Redmond, WA, USA) and Graphpad Prism 7.0. Red blood cell counts and percentages were reported to two significant figures. T-tests were used to compare blood cell counts and percentages between groups and at different time points within groups.

[0210] Experimental results:

[0211] (1) Effect on RBC count: The RBC count gradually increased after drug administration, and the 15mpk (mpk: mg / kg) drug administration group reached its highest point on the 29th day, and then stabilized. The hematocrit and hemoglobin concentration stabilized after the 29th day, and the drug administration group was higher than the control group, with an obvious dose-response relationship. Compared with before drug administration, the RBC counts of the 10 and 15mpk groups gradually increased on D15, D29, and D43, and were significantly dose-dependent. Although there was no significant difference between the 5mpk group and the pre-drug administration at each time point, the RBC count showed a slow upward trend; the RBC counts of the 10 and 15mpk groups increased significantly on D15, D29, and D43 compared with before drug administration, and the increase in the RBC count of the 15mpk group was more significant. See the table below:

[0212] (2) Effect on HGB hemoglobin content: The HGB content of each drug-treated group increased with the increase of drug administration time. Compared with the vehicle group, the 15mpk group had statistically significant differences on D29 and D43 (*P<0.05; ***P<0.001). The HGB content of the 10 and 15mpk groups on D15, D29, and D43 increased significantly, and showed obvious dose dependence, which was statistically significant compared with the pre-drug treatment. Compared with the pre-drug treatment, the HGB content of each group at each time point after drug administration was #, P<0.05; ##, P<0.01; ###, P<0.001; ####, P<0.0001. See the table below:

[0213] (3) Effect on HCT hematocrit: 5, 10, and 15 mpk of Comp-1 can increase the hematocrit HCT percentage. As the administration time increases, HCT gradually increases, and there is a certain degree of dose dependence. Compared with the vehicle group, at the end point of the experiment D43, the three dose groups were statistically significant (low dose group P < 0.05, medium dose group P < 0.01, high dose group P < 0.01). The effect of the high dose group in increasing HCT at D43 was significantly stronger than that of the medium and low dose groups (P < 0.05, P < 0.01). After administration, the low, medium, and high dose groups showed a significant increase in HCT% compared with their respective pre-administration results, and there was a clear dose dependence (#, P < 0.05; ##, P < 0.01; ###, P < 0.001; ####, P < 0.0001). See the table below:

[0214] It can be seen that Comp-1 can significantly increase the number of red blood cells, hemoglobin content, and hematocrit in thalassemia mice, indicating that Comp-1 has a therapeutic effect on anemia in homozygous d3th thalassemia genotype mice.

[0215] Example 5

[0216] Effects of HIF PH inhibitor and EPO on SCD anemia mice

[0217] Throughout the experimental period, animals were housed in cages with 24-hour central air conditioning, a temperature set at 20-26°C, a relative humidity of 40-70%, and a 12-hour light-dark cycle. All animals had free access to food and sterilized drinking water. The mice were B6.Cg-Hbbtm2(HBG1, HBB*)Tow / Hbatm1(HBA)Tow / ShmtJ mice, a Townse SCD model in which endogenous hemoglobin is replaced by humanized hemoglobin. Two weeks after birth, the mice were ear-punched and numbered. Ear tissue from the punched ears was collected and genotyped by PCR. Mice confirmed to be B6.Cg-Hbbtm2(HBG1, HBB*)Tow / Hbatm1(HBA)Tow / ShmtJ were grouped. When mice were approximately 4 weeks old, blood was collected from the eyes to measure RBC, Hb, and HCT values ​​to determine whether the animals showed signs of anemia. Blood samples were collected every other week. Mice showing signs of anemia were selected for drug administration. Animals were randomly divided into a vehicle control group (0.1 M meglumine aqueous solution; oral gavage; volume: 6.67 mL / kg), a Comp-1 group (30 mpk; oral gavage; volume: 6.67 mL / kg), and an EPO group (50 U / 30 g body weight; subcutaneous injection; MyBioSource, cat.#: MBS650094, erythropoietin alpha). Dosing was continued three times a week (TIW) for 4 consecutive weeks. On days 14 and 28 of drug administration, 70 μL of blood was collected from the eyes; the collected blood samples were placed in heparin-anticoagulant tubes. 15 μL of blood sample was drawn from the anticoagulant tube and loaded onto the Element HT5 Veterinary Hematology Analyzer. RBC, Hb, and HCT were measured according to the instrument's operating instructions. Data were analyzed using Graphpad Prism 9.0.

[0218] Experimental results:

[0219] From the arithmetic mean values ​​of various indicators (red blood cell count, hemoglobin content and hematocrit) of mice in each group before administration in the three tables below, it can be seen that the mice in each group were highly anemic, indicating that the model was successfully established.

[0220] (1) Effect on RBC count: As can be seen from the table below, the RBC count of the animals in the vehicle control group continued to decline over time, indicating that the degree of anemia was worsening. Even in the EPO-positive control mice, which can be used for anemia such as CKD, their RBC count continued to decline over time, and the degree of anemia was worsening. The RBC counts of the EPO group and the vehicle control group were close at the same time point, indicating that EPO could not induce an increase in RBC count. However, the RBC count of the Comp-1 group (30 mg / kg) increased significantly after 2 and 4 weeks of administration (8.88 M / μL, 9.09 M / μL vs 6.34 M / μL).

[0221] (2) Effect on HGB hemoglobin content: As can be seen from the table below, as time progresses, the hemoglobin content of the mice in the vehicle control group continues to decline, and the degree of anemia is deepening; even in the mice in the EPO positive control group, the hemoglobin content continues to decline, and the degree of anemia is deepening. The red blood cell counts of the EPO group and the vehicle control group at the same time point are close, indicating that EPO cannot induce an increase in hemoglobin content; while the hemoglobin content of the Comp-1 group (30 mg / kg) increased significantly after 2 and 4 weeks of administration (11.63 g / dL, 12.10 g / dL vs 8.47 g / dL), and was close to normal levels.

[0222] (3) Effect on HCT hematocrit: As can be seen from the table below, as time progressed, the hematocrit level of the mice in the vehicle control group continued to decline, and the degree of anemia continued to worsen; even for the mice in the EPO positive control group, the hematocrit level continued to decline, and the degree of anemia continued to worsen. The hematocrit levels of the EPO group and the vehicle control group were close at the same time point, indicating that EPO could not increase the hematocrit level; while the hematocrit level of the Comp-1 group (30 mg / kg) increased significantly after 2 and 4 weeks of administration compared with before administration (40.05%, 41.45% vs 27.20%), and was close to the normal level.

[0223] As controls, the same experiment was repeated using Roxadustat (30 mpk, 60 mkp, oral gavage, TIW / three times a week) and Daprodustat (30 mpk, oral gavage, TIW / three times a week).

[0224] At the end of the four-week experiment, both dose groups of Roxadustat failed to improve RBC, Hb, and HCT; a 2022 Nature article (Nautre 2022, vol 610, 783: https: / / www.nature.com / articles / s41586-022-05312-w) found that Roxadustat can induce fetal hemoglobin at the cellular level, indicating that Roxadustat may be used for SCD, but this experiment showed that Roxadustat's performance in SCD model mice was completely unexpected, and Roxadustat had no therapeutic effect at all on SCD mice.

[0225] Daprodustat can slightly improve RBC, Hb, and HCT. At the end of the four-week experiment, RBC increased from 6.14M / μL to 7.54M / μL, Hb increased from 9.1g / dL to 10.9g / dL, and HCT increased from 31.8% to 35.9%. One of the six mice in the Daprodustat group died two weeks after administration, so the baseline data of the experiment was the average of six mice, and the average of five mice at the end of four weeks. The results showed that Daprodustat had a slight effect on the SCD model mice, but was far inferior to Comp-1.

[0226] From the above effects, it can be seen that although EPO is known to be used for anemia such as CKD chronic kidney disease, it has no therapeutic effect in SCD model mice. Roxaudstat can induce fetal hemoglobin at the cellular level, but is completely ineffective in SCD model mice. Daprodustat can slightly improve RBC, Hb and HCT, but Comp-1 shows a surprising effect.

[0227] Although the embodiments of the present application have been illustrated and described, it is not intended that these embodiments illustrate and describe all possible forms of the present application. Rather, the words used in this specification are merely descriptive and not restrictive, and it should be understood that various changes can be made without departing from the spirit and scope of the present application.

Claims

1. Use of a hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) in the manufacture of a medicament for treating rare anemias.

2. Use of a hypoxia-inducible factor-proline hydroxylase inhibitor (HIF-PHI) in the manufacture of a medicament for increasing red blood cell count, hemoglobin content and / or hematocrit in rare anemias.

3. A method for treating rare anemia, comprising: A therapeutically effective amount of a hypoxia inducible factor-prolamide hydroxylase inhibitor (HIF-PHI) is administered to a patient in need of treatment.

4. A method for increasing the red blood cell count, hemoglobin content and / or hematocrit of a rare anemia, comprising: A therapeutically effective amount of a hypoxia inducible factor-prolamide hydroxylase inhibitor (HIF-PHI) is administered to a patient in need of treatment.

5. The use or method according to any one of claims 1 to 4, wherein: The rare anemia is selected from myelodysplastic syndrome anemia (MDS anemia), beta-thalassemia (β-thalassemia) or sickle cell disease (sickle cell anemia or SCD anemia).

6. The use or method of any one of claims 1 to 4, wherein the hypoxia inducible factor-prolyl hydroxylase inhibitor (HIF-PHI) is Comp-1.

7. The use or method according to any one of claims 1 to 4, wherein: The HIF-PHI is a compound of formula I or a pharmaceutically acceptable salt or a tautomer thereof: in: R 1 and R 2 are independently selected from cyano, alkyl, heterocyclic, alkenyl, alkynyl, aryl, wherein the above alkyl, heterocyclic, alkenyl, alkynyl, aryl, heteroaryl, acyl are optionally substituted by one or more substituents, wherein the substituents are independently selected from halogen, cyano, hydroxyl, amino, carboxyl, acyl, alkyl, heterocyclic, alkenyl, alkynyl, aryl, heteroaryl, =O, =S, -SH, R 10 O-、R 10 S-, R 10 (O=)S-、R 10 (O=)2S-, where R 10 is alkyl, heterocyclyl, alkenyl, alkynyl, aryl, or heteroaryl; or R 1 and R 2 Put together to form a ring; R 3 is selected from alkyl, heterocyclyl, alkenyl, alkynyl, aryl, and heteroaryl; R 4 and R 5 is hydrogen; R 6 and R 6’ is hydrogen; R 7 is hydrogen; R 8 is selected from hydrogen and alkyl; X is an oxygen atom.

8. The use or method according to claim 7, wherein in the compound of formula I, R 1 and R 2 Together with the C atoms to which they are attached, they form a cyclopentane ring (i.e., R 1 and R 2 is -CH2CH2CH2CH2-), R 3 is phenyl, R 4 and R 5 is hydrogen, R 6 and R 6’ is hydrogen, R 7 is hydrogen, R 8 is hydrogen, and X is an oxygen atom, that is, the compound of formula I is (6'-hydroxy-8'-oxo-3'-phenyl-8'H-spiro[cyclopentane-1,5'-indolizine]-7'-carbonyl)glycine.

9. The use or method according to any one of claims 1 to 4, wherein: When the rare anemia is myelodysplastic syndrome anemia (MDS anemia), the HIF-PHI is selected from: Daprodustat, Vadadustat, Molidustat, Enarodustat, Desidustat, HIF117 (SSS17), HEC53856, DDO-3055 and FG-2216.

10. The use or method according to any one of claims 1 to 4, wherein When the rare anemia is β-thalassemia (β-thalassemia), the HIF-PHI is selected from: roxadustat, Daprodustat, Vadadustat, Molidustat, Enarodustat, Desidustat, HIF117 (SSS17), HEC53856, DDO-3055 and FG-2216.

11. The use or method according to any one of claims 1 to 4, wherein: When the rare anemia is sickle cell disease (sickle cell anemia or SCD anemia), the HIF-PHI is selected from the group consisting of Daprodustat, Vadadustat, Molidustat, Enarodustat, Desidustat, HIF117 (SSS17), HEC53856 and DDO-3055.

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