Use of the PRDX5 and GPX4 genes in the manufacture of drugs for castration-resistant prostate cancer

Abstract

The present invention relates to the use of the PRDX5 and GPX4 genes in the manufacture of drugs for castration-resistant prostate cancer, and belongs to the field of biomedicine. The present invention proposes treating or supporting the treatment of castration-resistant prostate cancer by silencing or interfering with the expression of the PRDX5 and / or GPX4 genes. The present invention is the first to propose a strategy of combining siRNA with an androgen receptor antagonist to manufacture a drug for treating CRPC, and has conducted validation studies from multiple angles and dimensions. The present invention's pharmaceutical composition combining siRNA with an androgen receptor inhibitor is useful for the treatment of castration-resistant prostate cancer and effectively improves the inhibitory effect of enzalutamide on castration-resistant prostate cancer, laying the foundation for the design and clinical application of nucleic acid drugs and having important clinical therapeutic significance.

Description

[Technical Field]

[0001] The present invention relates to the field of biopharmaceuticals, and more specifically to the use of the genes PRDX5 and GPX4 in the manufacture of drugs for castration-resistant prostate cancer. [Background technology]

[0002] Androgen deprivation therapy is the standard treatment for advanced prostate cancer, but after an average of 1-3 years of treatment, patients eventually progress to castration-resistant prostate cancer (CRPC). CRPC refers to prostate cancer that progresses after initial, continuous androgen deprivation therapy (ADT). Since 2004, when docetaxel was proven to extend overall survival in patients with metastatic castration-resistant prostate cancer (mCRPC), drugs targeting the stage of mCRPC, such as abiraterone acetate, enzalutamide, and cabazitaxel, have been developed, changing the landscape of treatment for these patients. However, none of these have been able to completely reverse CRPC. Therefore, finding other effective therapeutic targets has become another hot spot in CRPC treatment research.

[0003] Peroxiredoxin 5 (PRDX5) is the only atypical 2-Cys peroxiredoxin in mammals, localizing in mitochondria, peroxisomes, cytoplasm, and the nucleus, and is a cytoprotective antioxidant enzyme against endogenous or exogenous peroxidative attack. Studies have shown that cancer cells require PRDX5 to maintain activity under oxidative stress (OS) conditions, and that overexpression of PRDX5 helps protect cells from apoptosis caused by OS. OS refers to a state of imbalance between oxidative and antioxidant activity in the body, resulting from an imbalance between two opposing forces: the generation of reactive oxygen species (ROS) and antioxidant activity. ROS can promote pathological defects in organisms such as cancer and diabetes, and can also damage cellular components such as DNA. ROS include hydrogen peroxide (H2O2), superoxide anion (O2O2), and superoxide anion (O2O2). 2- ROS are highly reactive oxygen species, including hydroxyl radicals. In cancer cells, ROS expression levels increase, but antioxidants such as PRDX5 present in cancer cells can detoxify ROS, thereby promoting the growth and development of cancer cells. When the balance between ROS and antioxidants between cells is disrupted, the intracellular ROS threshold level rises, and OS-mediated apoptosis occurs.

[0004] Phospholipid hydroperoxide glutathione peroxidase (GPX4) is an essential antioxidant peroxidase that directly reduces hydrogen peroxide phospholipids, even when incorporated into membranes and lipoproteins. It also reduces fatty acid hydroperoxides, cholesterol hydroperoxides, and thymine hydroperoxides. By preventing peroxidation of membrane lipids, it plays a crucial role in protecting cells from oxidative damage. A 2017 Nature report indicated that cancer cells with high interstitial therapy resistance are selectively susceptible to ferroptosis, a type of non-apoptotic cell death caused by iron-dependent accumulation of lipid reactive oxygen species, and that this can be induced by inhibiting lipid hydroperoxidase GPX4. Studies have shown that susceptibility to GPX4 inhibition is a common susceptibility in persistent cells.

[0005] Small molecule nucleic acid drugs exert their therapeutic effects through RNA interference (RNAi), selectively targeting corresponding targets, offering high specificity, and extending drug targets to upstream RNA of functional proteins to control target gene expression at the post-transcriptional level. In the early stages of siRNA therapy development, many drugs were designed based on fully unmodified or slightly modified siRNA to reach appropriate tissues and silence target genes. These molecules can mediate gene silencing in vivo. However, these methods have limited efficacy, and potential off-target effects may be observed. Therefore, chemically modified siRNA, such as substituting 2'-OH with 2'-O-methyl (2'-OMe) or 2'-methoxyethyl (2'-MOE) groups, can effectively inhibit innate immune activation by siRNA, enhance activity and specificity, and mitigate off-target induced toxicity. Many chemical modifications have been established and tested to enhance the potency of siRNA and reduce its potential toxicity. The present invention aims to slow the progression of CRPC by preparing PRDX5 and GPX4, which have been identified as effective therapeutic targets for CRPC, into modified siRNA drugs that exhibit RNA interference effects in cells and mice, thereby reducing the expression of PRDX5, GPX4, and PRDX5+GPX4. [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] The technical problem that this invention aims to solve is to discover an effective therapeutic target, provide siPRDX5, a nucleic acid drug that can effectively treat CRPC, and significantly improve the therapeutic effect of CRPC.

[0007] In this invention, extensive research and exploration have led to the discovery of nucleic acid drugs, namely siPRDX5 and siGPX4, for the treatment of CRPC. Research has shown that administration to human prostate cancer LNCaP cells, mouse MyC-CaP cells, and drug-tolerant persister (DTP) cells formed by LNCaP and MyC-CaP cells reduces the expression of PRDX5 and siGPX4 in cancer cells with either siPRDX5 or siGPX4 alone, while the combination of siPRDX5 and siGPX4 reduces the expression of both PRDX5 and GPX4. Gene and protein expression were verified by q-RTPCR and Western Blot assays.

[0008] In this invention, LNCaP / MyC-CaP-DTP cells and 22Rv1 cells were treated with siPRDX5, siGPX4, or siPRDX5+siGPX4, and the effects of the drugs were verified through CCK8 cell proliferation analysis. The results showed that siPRDX5, siGPX4, or siPRDX5+siGPX4 significantly inhibited the growth of LNCaP-DTP, MyC-CaP-DTP, and 22Rv1 cells. Furthermore, a CRPC mouse model was constructed by administering ENZ sequentially intragastricly to C-MYC overexpressing prostate cancer mice, and siPRDX5, siGPX4, or siPRDX5+siGPX4 were administered to the CRPC mice. The enzymatic activity of PRDX5 in the mice was examined to confirm the therapeutic effect of this drug on prostate cancer. The results indicate that siPRDX5, siGPX4, or siPRDX5+siGPX4 effectively inhibit the enzymatic activity of PRDX5, thereby effectively inhibiting the progression of prostate cancer in CRPC mice.

[0009] The first object of the present invention is to provide a double-stranded siRNA molecule as shown in any of (A) to (C) below.

[0010] (A) An siRNA molecule that inhibits PRDX5 gene expression, having a double-stranded siRNA molecule in which the single-stranded RNA indicated by SEQ ID NO.1 and the single-stranded RNA indicated by SEQ ID NO.2 are complementary. (B) siRNA molecules that inhibit GPX4 gene expression, comprising a double-stranded siRNA molecule in which the RNA single strand indicated by SEQ ID NO.3 and the RNA single strand indicated by SEQ ID NO.4 are complementary, or an siRNA molecule in which the RNA single strand indicated by SEQ ID NO.5 and the RNA single strand indicated by SEQ ID NO.6 are complementary, and A composition containing (C)(A) and (B).

[0011] In one embodiment of the present invention, at least one nucleotide in the double-stranded siRNA molecule is a modified nucleotide.

[0012] In one embodiment of the present invention, all nucleotides in the double-stranded siRNA molecule are modified nucleotides.

[0013] In one embodiment of the present invention, the modifications are phosphorothioate, 2'-F, 2'-OMe, 2'-Ara-F, 2'-O-MOE, m 6 A or m 5 It is at least one selected from C.

[0014] In one embodiment of the present invention, the modification is shown in any of (1) to (7) below. TIFF0007917934000001.tif47170(1) Phosphothioate (PS, Rp isomer)

[0015] TIFF0007917934000002.tif45170(2)2'-deoxy-2'-fluoro(2'-F)

[0016] TIFF0007917934000003.tif45170(3)2'-O-methyl(2'-OMe)

[0017] TIFF0007917934000004.tif45170(4)2'-Arabino-fluoro (2'-Ara-F)

[0018] TIFF0007917934000005.tif49170(5)2'-O-methoxymethyl (2'-O-MOE)

[0019] TIFF0007917934000006.tif58170(6)N6'-methyladenosine (m 6 A)

[0020] TIFF0007917934000007.tif59170(7)5'-methylcytidine (m 5 C)

[0021] In one embodiment of the present invention, the double-stranded siRNA molecule is shown in any one of (A1)~(A16), (B1)~(B16), and (C1)~(C16), wherein (A1)~(A16) are modified double-stranded siRNAs targeting siPRDX5, and (B1)~(B16) and (C1)~(C16) are modified double-stranded siRNAs targeting siGPX4.

[0022] (A1): 5' (C)-[A]-(G)[A](C)[U](U)[A](UUA)[C](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C)[A](U)[C](U)[A](G)[UAA](U)[A](A)[G](U)[C](U)[G]-[U]-[C] 3'; (A2) 5' (C)- A -(G) A (C) U (U) A (UUA) C (U) A (G) A (U) G (A) U (T) 3'; 5'A ]-(A)-[ U ](C)[ A ](U)[ C ](U)[ A ](G)[ UAA ](U)[ A ](A)[ G ](U)[ C ](U)[ G ]-[ U ]-[ C ] 3’; (A3) 5’ ( C )-[A]-( G )[A]( C )[U]( U )[A]( UUA )[C]( U )[A]( G )[A]( U )[G]( A )[U]( T ) 3’; 5’ [A]-( A )-[U]( C )[A]( U )[C]( U )[A]( G )[UAA]( U )[A]( A )[G]( U )[C]( U )[G]-[U]-[C] 3’; (A4) 5’ ( C )-[ A ]-( G )[ A ]( C )[ U ]( U )[ A ]( UUA )[ C ]( U )[ A ]( G )[ A ]( U )[ G ]( A )[ U ]( T ) 3’; 5’ [ A ]-( A )-[U ]( C )[ A ]( U )[ C ]( U )[ A ]( G )[ UAA ]( U )[ A ]( A )[ G ]( U )[ C ]( U )[ G ]-[ U ]-[ C ] 3'; (A5) 5' (C)-[A*]-(G)[A*](C)[U](U)[A*](UUA*)[C](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5' [A*]-(A*)-[U](C)[A*](U)[C](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C](U)[G]-[U]-[C] 3'; (A6) 5' (C')-[A]-(G)[A](C')[U](U)[A](UUA)[C'](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C')[A](U)[C'](U)[A](G)[UAA](U)[A](A)[G](U)[C'](U)[G]-[U]-[C'] 3'; (A7) 5' (C)-[ A *]-(G)[ A *](C)[ U ](U)[ A *](US*)[ C ](U)[ A *](G)[ A *](U)[ G ](A*)[ U ](T) 3'; 5'[ A *]-(A*)-[ U ](C)[ A *](U)[ C ](U)[ A*](G)[ UA * A *](U)[ A *](A*)[ G ](U)[ C ](U)[ G ]-[ U ]-[ C ] 3’; (A8) 5’ ( C )-[A*]-( G )[A*]( C )[U]( U )[A*]( UUA *)[C]( U )[A*]( G )[A*]( U )[G]( A *)[U]( T ) 3’; 5’ [A*]-( A *)-[U]( C )[A*]( U )[C]( U )[A*]( G )[UA*A*]( U )[A*]( A *)[G]( U )[C]( U )[G]-[U]-[C] 3’; (A9) 5’ ( C )-[ A *]-( G )[ A *]( C )[ U ]( U )[ A *]( UUA *)[ C ]( U )[ A *]( G )[ A *]( U )[ G ]( A *)[ U ]( T ) 3’; 5’ [ A *]-( A *)-[ U ]( C )[ A *](U )[ C ]( U )[ A *]( G )[ UA * A *]( U )[ A *]( A *)[ G ]( U )[ C ]( U )[ G ]-[ U ]-[ C ] 3’; (A10) 5’ (C’)-[ A ]-(G)[ A ](C’)[ U ](U)[ A ](UUA)[ C ’](U)[ A ](G)[ A ](U)[ G ](A)[ U ](T) 3’; 5’ [ A ]-(A)-[ U ](C’)[ A ](U)[ C ’](U)[ A ](G)[ UAA ](U)[ A ](A)[ G ](U)[ C ’](U)[ G ]-[ U ]-[ C ’] 3’; (A11) 5’ ( C ’)-[A]-( G )[A]( C ’)[U]( U )[A]( UUA )[C’]( U )[A]( G )[A]( U )[G]( A )[U]( T ) 3’; 5’ [A]-( A )-[U]( C ’)[A]( U)[C’]( U )[A]( G )[UAA]( U )[A]( A )[G]( U )[C’]( U )[G]-[U]-[C’] 3’; (A12) 5’ ( C ’)-[ A ]-( G )[ A ]( C ’)[ U ]( U )[ A ]( UUA )[ C ’]( U )[ A ]( G )[ A ]( U )[ G ]( A )[ U ]( T ) 3’; 5’ [ A ]-( A )-[ U ]( C ’)[ A ]( U )[ C ’]( U )[ A ]( G )[ UAA ]( U )[ A ]( A )[ G ]( U )[ C ’]( U )[ G ]-[ U ]-[ C ’] 3’; (A13) 5’ (C’)-[A*]-(G)[A*](C’)[U](U)[A*](UUA*)[C’](U)[A*](G)[A*](U)[G](A*)[U](T) 3’; 5’[A*]-(A*)-[U](C’)[A*](U)[C’](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C’](U)[G]-[U]-[C’]3’; (A14) 5’ (C’)-[ A *]-(G)[ A *](C’)[ U ](U)[ A *](UUA*)[ C ’](U)[ A *](G)[ A *](U)[ G ](A*)[ U ](T) 3’; 5’[ A *]-(A*)-[ U ](C’)[ A *](U)[ C ’](U)[ A *](G)[ UA * A *](U)[ A *](A*)[ G ](U)[ C ’](U)[ G ]-[ U ]-[ C ’]3’; (A15) 5’ ( C ’)-[A*]-( G )[A*]( C ’)[U]( U )[A*]( UUA *)[C’]( U )[A*]( G )[A*]( U )[G]( A *)[U]( T ) 3’; 5’[A*]-( A *)-[U]( C ’)[A*]( U )[C’]( U )[A*]( G )[UA*A*]( U )[A*]( A *)[G]( U )[C’]( U )[G]-[U]-[C’]3’; (A16) 5' ( C ')-[ A *]-( G )[ A *]( C ')[ U ]( U )[ A *]( UUA *)[ C ']( U )[ A *]( G )[ A *]( U )[ G ]( A *)[ U ]( T ) 3'; 5'[ A *]-( A *)–[ U ]( C ')[ A *]( U )[ C ']( U )[ A *]( G )[ UA * A *]( U )[ A *]( A *)[ G ]( U )[ C ']( U )[ G ]–[ U ]–[ C ']3'; (B1) 5' (A)-[C]-(A)[A](C)[G](U)[C](AAA)[U](U)[C](G)[A](U)[A](U)[G](U) 3'; 5' [A]-(C)-[A](U)[A](U)[C](G)[A](A)[UUU](G)[A](C)[G](U)[U](G)[U]-[A]-[C] 3'; (B2) 5′ (A)–[ C ]-(YOUR)[ A ](C)[ G ](HER)[ C ](AAA)[U ](THE)[ C ](G)[ A ](THE)[ A ](THE)[ G ](U) 3'; 5' [ A ]-(C)-[ A ](THE)[ A ](THE)[ C ](G)[ A ](WITH)[ UUU ](G)[ A ](C)[ G ](THE)[ U ](G)[ U ]-[ A ]-[ C ] 3'; (B3) 5' ( A )-[C]-( A )[WITH]( C )[G]( U )[C]( AAA )[THE]( U )[C]( G )[WITH]( U )[WITH]( U )[G]( U ) 3'; 5' [A]-( C )-[WITH]( U )[WITH]( U )[C]( G )[WITH]( A )[UUU]( G )[WITH]( C )[G]( U )[THE]( G )[U]-[A]-[C] 3'; (B4) 5' ( A )-[ C ]-( A )[ A ]( C )[ G ]( U )[ C ]( AAA )[ U ]( U )[ C ]( G )[ A ]( U )[A ]( U )[ G ]( U ) 3’; 5’ [ A ]-( C )-[ A ]( U )[ A ]( U )[ C ]( G )[ A ]( A )[ UUU ]( G )[ A ]( C )[ G ]( U )[ U ]( G )[ U ]-[ A ]-[ C ] 3’; (B5) 5’ (A*)-[C]-(A*)[A*](C)[G](U)[C](A*A*A*)[U](U)[C](G)[A*](U)[A*](U)[G](U) 3’; 5’ [A*]-(C)-[A*](U)[A*](U)[C](G)[A*](A*)[UUU](G)[A*](C)[G](U)[U](G)[U]-[A*]-[C] 3’; (B6) 5’ (A)-[C’]-(A)[A](C’)[G](U)[C’](AAA)[U](U)[C’](G)[A](U)[A](U)[G](U) 3’; 5’ [A]-(C’)-[A](U)[A](U)[C’](G)[A](A)[UUU](G)[A](C’)[G](U)[U](G)[U]-[A]-[C’] 3’; (B7) 5’ (A*)-[ C ]-(A*)[ A *](C)[ G ](U)[ C ](A*A*A*)[ U ](U)[ C ](G)[ A *](U)[ A *](U)[ G](U) 3'; 5' [ A *]-(W)-[ A *](U)[ A *](U)[ C ](G)[ A *](THE*)[ UUU ](G)[ A *](W)[ G ](U)[ U ](G)[ U ]-[ A *]-[ C ] 3'; (B8) 5' ( A *)-[W]-( A *)[THE*]( C )[G]( U )[W]( A * A * A *)[U]( U )[W]( G )[THE*]( U )[THE*]( U )[G]( U ) 3'; 5' [A*]-( C )-[THE*]( U )[THE*]( U )[W]( G )[THE*]( A *)[UUU]( G )[THE*]( C )[G]( U )[U]( G )[U]-[A*]-[C] 3'; (B9) 5' ( A *)-[ C ]-( A *)[ A *]( C )[ G ]( U )[ C ]( A * A * A *)[ U ]( U )[ C ]( G )[ A *]( U )[A *]( U )[ G ]( U ) 3’; 5’ [ A *]-( C )-[ A *]( U )[ A *]( U )[ C ]( G )[ A *]( A *)[ UUU ]( G )[ A *]( C )[ G ]( U )[ U ]( G )[ U ]-[ A *]-[ C ] 3’; (B10) 5’ (A)-[ C ’]-(A)[ A ](C’)[ G ](U)[ C ’](AAA)[ U ](U)[ C ’](G)[ A ](U)[ A ](U)[ G ](U) 3’; 5’ [ A ]-(C’)-[ A ](U)[ A ](U)[ C ’](G)[ A ](A)[ UUU ](G)[ A ](C’)[ G ](U)[ U ](G)[ U ]-[ A ]-[ C ’] 3’; (B11) 5’ ( A )-[C’]-( A )[A]( C ’)[G]( U )[C’]( AAA )[U]( U)[C’]( G )[A]( U )[A]( U )[G]( U ) 3’; 5’ [A]-( C ’)-[A]( U )[A]( U )[C’]( G )[A]( A )[UUU]( G )[A]( C ’)[G]( U )[U]( G )[U]-[A]-[C’] 3’; (B12) 5’ ( A )-[ C ’]-( A )[ A ]( C ’)[ G ]( U )[ C ’]( AAA )[ U ]( U )[ C ’]( G )[ A ]( U )[ A ]( U )[ G ]( U ) 3’; 5’ [ A ]-( C ’)-[ A ]( U )[ A ]( U )[ C ’]( G )[ A ]( A )[ UUU ]( G )[ A ]( C ’)[ G ]( U )[ U ]( G )[ U ]-[ A ]-[ C ’] 3’; (B13) 5’ (A*)-[C’]-(A*)[A*](C’)[G](U)[C’](A*A*A*)[U](U)[C’](G)[A*](U)[A*](U)[G](U) 3’; 5’ [A*]-(C’)-[A*](U)[A*](U)[C’](G)[A*](A*)[UUU](G)[A*](C’)[G](U)[U](G)[U]-[A*]-[C’] 3’; (B14) 5’ (A*)-[ C ’]-(A*)[ A *](C’)[ G ](U)[ C ’](A*A*A*)[ U ](U)[ C ’](G)[ A *](U)[ A *](U)[ G ](U) 3’; 5’ [ A *]-(C’)-[ A *](U)[ A *](U)[ C ’](G)[ A *](A*)[ UUU ](G)[ A *](C’)[ G ](U)[ U ](G)[ U ]-[ A *]-[ C ’] 3’; (B15) 5’ ( A *)-[C’]-( A *)[A*]( C ’)[G]( U )[C’]( A * A * A *)[U]( U )[C’]( G )[A*]( U )[A*]( U )[G]( U ) 3’; 5’ [A*]-( C ’)-[A*]( U )[A*]( U )[C’]( G )[A*](A *)[UUU]( G )[A*]( C ')[G]( U )[U]( G )[U]-[A*]-[C'] 3'; (B16) 5' ( A *)-[ C ']-( A *)[ A *]( C ')[ G ]( U )[ C ']( A * A * A *)[ U ]( U )[ C ']( G )[ A *]( U )[ A *]( U )[ G ]( U ) 3'; 5' [ A *]-( C ')-[ A *]( U )[ A *]( U )[ C ']( G )[ A *]( A *)[ UUU ]( G )[ A *]( C ')[ G ]( U )[ U ]( G )[ U ]-[ A *]-[ C '] 3'; (C1) 5' (G)-[U]-(G)[A](G)[G](C)[A](AGA)[C](C)[G](A)[A](G)[U](A)[A](A) 3'; 5’ [U]-(U)-[U](A)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A)[C]-[U]-[G] 3’; (C2) 5’ (G)-[ U ]-(G)[ A ](G)[ G ](C)[ A ](AGA)[ C ](C)[ G ](A)[ A ](G)[ U ](A)[ A ](A) 3’; 5’ [ U ]-(U)-[ U ](A)[ C ](U)[ U ](C)[ G ](G)[ UCU ](U)[ G ](C)[ C ](U)[ C ](A)[ C ]-[ U ]-[ G ] 3’; (C3) 5’ ( G )-[U]-( G )[A]( G )[G]( C )[A]( AGA )[C]( C )[G]( A )[A]( G )[U]( A )[A]( A ) 3’; 5’ [U]-( U )-[U]( A )[C]( U )[U]( C )[G]( G )[UCU]( U )[G]( C )[C]( U )[C]( A )[C]-[U]-[G] 3’; (C4) 5’ ( G )-[ U ]-(G )[ A ]( G )[ G ]( C )[ A ]( AGA )[ C ]( C )[ G ]( A )[ A ]( G )[ U ]( A )[ A ]( A ) 3’; 5’ [ U ]-( U )-[ U ]( A )[ C ]( U )[ U ]( C )[ G ]( G )[ UCU ]( U )[ G ]( C )[ C ]( U )[ C ]( A )[ C ]-[ U ]-[ G ] 3’; (C5) 5’ (G)-[U]-(G)[A*](G)[G](C)[A*](A*GA*)[C](C)[G](A*)[A*](G)[U](A*)[A*](A*) 3’; 5’ [U]-(U)-[U](A*)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A*)[C]-[U]-[G] 3’; (C6) 5’ (G)-[U]-(G)[A](G)[G](C’)[A](AGA)[C’](C’)[G](A)[A](G)[U](A)[A](A) 3’; 5’ [U]-(U)-[U](A)[C’](U)[U](C’)[G](G)[UC’U](U)[G](C’)[C’](U)[C’](A)[C’]-[U]-[G] 3’; (C7) 5’ (G)-[ U ]-(G)[ A *](G)[ G ](C)[ A *](A*GA*)[ C ](C)[ G ](A*)[ A *](G)[ U ](A*)[ A *](A*) 3’; 5’ [ U ]-(U)-[ U ](A*)[ C ](U)[ U ](C)[ G ](G)[ UCU ](U)[ G ](C)[ C ](U)[ C ](A*)[ C ]-[ U ]-[ G ] 3’; (C8) 5’ ( G )-[U]-( G )[A*]( G )[G]( C )[A*]( A * GA *)[C]( C )[G]( A *)[A*]( G )[U]( A *)[A*]( A *) 3’; 5’ [U]-( U )-[U]( A *)[C]( U )[U]( C )[G]( G )[UCU]( U )[G]( C )[C]( U )[C]( A *)[C]-[U]-[G] 3’; (C9) 5’ ( G )-[ U ]-( G )[ A *]( G )[ G ](C )[ A *]( A * GA *)[ C ]( C )[ G ]( A *)[ A *]( G )[ U ]( A *)[ A *]( A *) 3’; 5’ [ U ]-( U )-[ U ]( A *)[ C ]( U )[ U ]( C )[ G ]( G )[ UCU ]( U )[ G ]( C )[ C ]( U )[ C ]( A *)[ C ]-[ U ]-[ G ] 3’; (C10) 5’ (G)-[ U ]-(G)[ A ](G)[ G ](C’)[ A ](AGA)[ C ’](C’)[ G ](A)[ A ](G)[ U ](A)[ A ](A) 3’; 5’ [ U ]-(U)-[ U ](A)[ C ’](U)[ U ](C’)[ G ](G)[ UC ’ U ](U)[ G ](C’)[ C ’](U)[ C ’](A)[ C ’]-[ U ]-[G ] 3’; (C11) 5’ ( G )-[U]-( G )[A]( G )[G]( C ’)[A]( AGA )[C’]( C ’)[G]( A )[A]( G )[U]( A )[A]( A ) 3’; 5’ [U]-( U )-[U]( A )[C’]( U )[U]( C ’)[G]( G )[UC’U]( U )[G]( C ’)[C’]( U )[C’]( A )[C’]-[U]-[G] 3’; (C12) 5’ ( G )-[ U ]-( G )[ A ]( G )[ G ]( C ’)[ A ]( AGA )[ C ’]( C ’)[ G ]( A )[ A ]( G )[ U ]( A )[ A ]( A ) 3’; 5’ [ U ]-( U )-[ U ]( A )[ C ’]( U )[ U ]( C ’)[ G ]( G )[ UC ’ U ]( U )[ G ](C ’)[ C ’]( U )[ C ’]( A )[ C ’]-[ U ]-[ G ] 3’; (C13) 5’ (G)-[U]-(G)[A*](G)[G](C’)[A*](A*GA*)[C’](C’)[G](A*)[A*](G)[U](A*)[A*](A*) 3’; 5’ [U]-(U)-[U](A*)[C’](U)[U](C’)[G](G)[UC’U](U)[G](C’)[C’](U)[C’](A*)[C’]-[U]-[G] 3’; (C14) 5’ (G)-[ U ]-(G)[ A *](G)[ G ](C’)[ A *](A*GA*)[ C ’](C’)[ G ](A*)[ A *](G)[ U ](A*)[ A *](A*) 3’; 5’ [ U ]-(U)-[ U ](A*)[ C ’](U)[ U ](C’)[ G ](G)[ UC ’ U ](U)[ G ](C’)[ C ’](U)[ C ’](A*)[ C ’]-[ U ]-[ G ] 3’; (C15) 5’ ( G )-[U]-( G )[A*]( G )[G]( C ’)[A*]( A * GA *)[C’]( C ’)[G]( A *)[A*](G )[U]( A *)[A*]( A *) 3'; 5' [U]-( U )-[U]( A *)[C']( U )[U]( C ')[G]( G )[UC'U]( U )[G]( C '[C']( U )[C']( A *)[C']-[U]-[G] 3'; (C16) 5' ( G )-[ U ]-( G )[ A *]( G )[ G ]( C ')[ A *]( A * GA *)[ C ']( C ')[ G ]( A *)[ A *]( G )[ U ]( A *)[ A *]( A *) 3'; 5' [ U ]-( U )-[ U ]( A *)[ C ']( U )[ U ]( C ')[ G ]( G )[ UC ' U ]( U )[ G ]( C ')[ C ']( U )[ C ']( A *)[ C ']-[ U ]-[ G ] 3'; (Here, A-, U-, C-, and G- represent phosphorothioate-modified ribonucleotides A, U, C, and G, respectively.) (A), (U), (C), and (G) represent ribonucleotides A, U, C, and G, respectively, that have undergone 2'-F modification. [A], [U], [C], and [G] represent ribonucleotides A, U, C, and G, respectively, that have undergone 2'-OMe modification. (A) , (U) , (C) , and (G) These represent ribonucleotides A, U, C, and G, respectively, which have undergone 2'-Ara-F modification. [A] , [U] , [C] , and [G] These represent ribonucleotides A, U, C, and G, respectively, which have undergone 2'-O-MOE modification. A*, U*, C*, and G* are each m 6 This shows ribonucleotides A, U, C, and G that have undergone A modification. A', U', C', and G' are each m 5 (This shows ribonucleotides A, U, C, and G that have undergone C modification.)

[0023] The second object of the present invention is (1) A vector expressing the siRNA, (2) Host cells having the siRNA or the vector, (3) Reagents containing the siRNA, or (1) the vector or (2) the host cell, and (4) Pharmaceutical composition containing the siRNA The objective is to provide an siRNA-related biomaterial that is one of the following.

[0024] In one embodiment of the present invention, the pharmaceutical composition further comprises an androgen or an androgen receptor inhibitor.

[0025] In one embodiment of the present invention, the androgen receptor inhibitor is selected from enzalutamide (ENZ).

[0026] In one embodiment of the present invention, the pharmaceutical composition contains a pharmaceutically acceptable carrier or auxiliary material.

[0027] In one embodiment of the present invention, the pharmaceutically acceptable carrier includes microliposomes, microcells, metal particles, or polymer particles.

[0028] A third object of the present invention is to provide the use of the siRNA or the biomaterial in the manufacture of a drug for preventing or treating prostate cancer.

[0029] In one embodiment of the present invention, the prostate cancer is advanced prostate cancer or castration-resistant prostate cancer.

[0030] A fourth object of the present invention is to provide a method for preventing or treating prostate cancer by administering the siRNA or the biological material to a subject.

[0031] A fifth object of the present invention is to provide a method for inhibiting the expression of PRDX5 and / or GPX4 genes in cells, comprising contacting an effective amount of the siRNA or the biological material with cells to inhibit the expression of PRDX5 and / or GPX4 genes in cells. [Effects of the Invention]

[0032] Beneficial effects 1. The present invention proposes that by interfering with or silencing the expression of the genes PRDX5 and / or GPX4, prostate cancer, particularly castration-resistant prostate cancer, can be improved, exhibiting a clear killing effect on castration-resistant prostate cancer cells with a maximum cell inhibition rate of approximately 50%, reducing the prostate weight of drug-resistant CRPC mice, suppressing tumor growth, and, when used in combination with the androgen receptor inhibitor ENZ, reducing the prostate weight to approximately 40 mg, and also significantly inhibiting PRDX5 enzyme activity in the prostate.

[0033] 2. This invention is the first to propose the production of a CRPC treatment drug using the nucleic acid drugs siPRDX5 and / or siPGX4, and is of great significance in promoting the application of nucleic acid drugs in the clinical treatment of prostate cancer. Drug research takes an average of 8 to 10 years from compound molecules to clinical applications, requiring many human resources and material support, and incurring enormous time and economic costs. The solution of this invention can significantly shorten the time from drug discovery to clinical application. [Brief explanation of the drawing]

[0034] Figure 1 This describes the detection of PRDX5 and GPX4m RNA expression levels by qRT-PCR after adding siRNAs #1, #2, and #3siPRDX5, and #1, #2, #3, and #4siGPX4 according to the present invention to LNCaP and LNCaP-derived DTP cells (LNCaP-DTP). Figure A shows the relative expression level of PRDX5 in LNCaP cells, Figure B shows the relative expression level of PRDX5 in LNCaP-DTP cells, Figure C shows the relative expression level of GPX4 in LNCaP cells, and Figure D shows the relative expression level of GPX4 in LNCaP-DTP cells. Figure 2This study involves detecting the mRNA expression level of PRDX5 by qRT-PCR after adding the modified #3siPRDX5 according to the present invention to LNCaP cells, MyC-CaP cells, LNCaP-derived DTP cells (LNCaP-DTP), and MyC-CaP-derived DTP cells (MyC-CaP-DTP). Figure A shows LNCaP cells, Figure B shows MyC-CaP cells, Figure C shows LNCaP-DTP cells, and Figure D shows MyC-CaP-DTP cells. Figure 3 This involves detecting the mRNA expression levels of GPX4 by qRT-PCR after adding modified #3 and #4siGPX4 according to the present invention to LNCaP cells, MyC-CaP cells, LNCaP-derived DTP cells (LNCaP-DTP), and MyC-CaP-derived DTP cells (MyC-CaP-DTP). Figure A shows LNCaP cells, Figure B shows MyC-CaP cells, Figure C shows LNCaP-DTP cells, and Figure D shows MyC-CaP-DTP cells. Figure 4 This study involved detecting changes in PRDX5 or GPX4 protein expression by Western blotting after adding unmodified #3siPRDX5 or siGPX4 to LNCaP cells and MyC-CaP cells. Figure A shows the detection of PRDX5 protein expression in LNCaP cells, Figure B shows the detection of PRDX5 protein expression in MyC-CaP cells, Figure C shows the detection of GPX4 protein expression in LNCaP cells, and Figure D shows the detection of GPX4 protein expression in MyC-CaP cells. Figure 5This study detects cell viability by CCK8 after adding modified #3siPRDX5-1-16, modified #3siGPX4-1-16, or modified #3siPRDX5-1-16 + modified #3siGPX4-1-16 to LNCaP-DTP cells and MyC-CaP-DTP cells. Figure A shows the effect of siPRDX5-1-16 on the viability of LNCaP-DTP cells, Figure B shows the effect of siPRDX5-1-16 on the viability of MyC-CaP-DTP cells, Figure C shows the effect of siPRDX5-1-16 on the viability of 22Rv1 cells, Figure D shows the effect of siGPX4-1-16 on the viability of LNCaP-DTP cells, and Figure E shows the effect of siGPX4-1-16 on the viability of MyC-CaP-DTP cells. Figure F shows the effect on survival rates of 22Rv1 cells due to the addition of siGPX4-1-16, Figure G shows the effect on survival rates of LNCaP-DTP cells due to the addition of siPRDX5-1-16 + siGPX4-1-16, Figure H shows the effect on survival rates of MyC-CaP-DTP cells due to the addition of siPRDX5-1-16 + siGPX4-1-16, and Figure I shows the effect on survival rates of 22Rv1 cells due to the addition of siPRDX5-1-16 + siGPX4-1-16. Figure 6 Figure A shows the effect of the following on changes in prostate weight in C-MYC overexpressing prostate cancer mice in which CRPC was induced by continuous administration of ENZ: NC control group, ENZ, #3siPRDX5-1, #3siPRDX5-16, #3siGPX4-1, #3siGPX4-16, #3siPRDX5-16 + #3siGPX4-16, and combination therapy with ENZ. Figure A shows the change in body weight of mice in each group (NC, ENZ, siPRDX5-1, ENZ + siPRDX5-1 group) as administration progresses, Figure B is a comparative image of photographs of prostates removed from 8-month-old mice in each group, and Figure C shows the change in prostate weight of 8-month-old mice in each group. Figure 7This figure shows the changes in PRDX5 protease activity in C-MYC overexpressing prostate cancer mice in which CRPC was induced by continuous administration of ENZ, in the NC control group, ENZ, #3siPRDX5-1, #3siPRDX5-16, #3siGPX4-1, #3siGPX4-16, #3siPRDX5-16 + #3siGPX4-16, and in combination with ENZ. Figure A shows HE staining of prostate tissue sections from 8-month-old mice of each group, and Figure B shows the quantitative PRDX5 enzyme activity in the prostate tissue of mice of each group. [Modes for carrying out the invention]

[0035] The present invention will be further described below with reference to the drawings and specific examples, but these examples are not intended to limit the present invention in any way. Unless otherwise specified, the reagents, methods and apparatus used in the present invention are conventional reagents, methods and apparatus in the art.

[0036] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0037] Example 1 Synthesis of unmodified siRNA Based on the complete mRNA sequences of human PRDX5 and GPX4, siRNAs were designed, and the activity of all candidate siRNAs was evaluated through the basic design principles of siRNA. The siRNAs were then designed and synthesized. All sequences were obtained from the NCBI gene database.

[0038] (1) siPRDX5 sequence: #1: 5'-AGACAGACUUAUUACUAGAUGAUTC-3'(SEQ ID NO.7) 5'-GAAUCAUCUAGUAAUAAGUCUGUCUCC-3'(SEQ ID NO.8) #2: 5' GGCCAGAUUUCUGCAAUAAACACTT-3'(SEQ ID NO.9) 5' AAGUGUUUAUUGCAGAAAUCUGGCCAA-3'(SEQ ID NO.10) #3: 5'-CAGACUUAUUACUAGAUGAUT-3'(SEQ ID NO.1) 5'-AAUCAUCUAGUAAUAAAGUCUGUC-3'(SEQ ID NO.2) (2) siGPX4 array: #1: 5'-GUGAGGCAAGACCGAAGUAAACUAC- 3'(SEQ ID NO.11) 5'-GUAGUUUACUUCGGUCUUGCCUCACUG-3'(SEQ ID NO.12) #2: 5'-CUACAACGUCAAAUUCGAUAUGUTC-3'(SEQ ID NO.13) 5'-GAACAUAUCGAAUUUGACGUUGUAGCC-3'(SEQ ID NO.14) #3: 5'-ACAACGUCAAAUUCGAUAUGU-3'(SEQ ID NO.3) 5'-ACAUAUCGAAUUUGACGUUGUAC-3'(SEQ ID NO.4) #4: 5'-GUGAGGCAAGACCGAAGUAAA-3'(SEQ ID NO.5) 5'-UUUACUUCGGUCUUGCCUCACUG-3'(SEQ ID NO.6) Example 2 Gene interference ability of unmodified siPRDX5 and siGPX4 on prostate cancer cells and DTP cells To screen for suitable siRNAs, the interference activity of the siRNAs designed in Example 1 was detected.

[0039] S1. Place LNCaP and MyC-CaP cells in a 10cm cell culture dish (1 × 10⁶). 6Individual seeds were seeded, and after attachment the following day, 50 μM enzalutamide (ENZ) was added and the cells were treated for 9 days. During this time, the culture medium was changed to fresh drug-containing medium (1640 medium containing 50 μM ENZ) every 3 days, and the cells were collected. The cells treated for 9 days were designated as drug-resistant cells (DTP cells), and were named LNCaP-DTP cells and MyC-CaP-DTP cells, respectively.

[0040] S2.LNCaP cells, MyC-CaP cells, and LNCaP-DTP cells and MyC-CaP-DTP cells collected in S1 were inoculated into 6cm and 10cm dishes containing fresh complete medium (1640 medium containing 10% fetal bovine serum and 1% biantibody). After adhesion, the cells were transfected with siPRDX5 and siGPX4 designed in Example 1, and then mRNA and protein were extracted.

[0041] S3.qRT-PCR and Western Blot testing were used to verify at the RNA and protein levels that siRNAs with interference activity were screened for.

[0042] As shown in Figures 1 and 4, and Tables 1 and 2, the addition of siPRDX5 #3siRNA significantly reduced PRDX5 expression, including mRNA and protein levels, while the addition of siGPX4 #3 and #4 siRNAs drastically reduced GPX4 expression, including mRNA and protein levels. In human prostate cancer cells (LNCaP cells) and drug-resistant LNCaP-DTP cells, PRDX5 mRNA expression levels decreased to approximately 0.16 after siPRDX5 #3 siRNA interference, but PRDX5 mRNA expression levels only decreased to approximately 0.16 after siPRDX5 #1 and #2 siRNA interference. In human prostate cancer cells (LNCaP) and drug-resistant LNCaP-DTP cells, the mRNA expression level of GPX4 decreased to approximately 0.21 after interference with siGPX4 using #3 siRNA, and decreased to approximately 0.24 after interference with siGPX4 using #4 siRNA. On the other hand, the mRNA expression level of GPX4 decreased to approximately 0.73 after interference with siGPX4 using #1 and #2 siRNA.

[0043] Based on the above results, the siRNA designed in Example 1 of the present invention has the ability to interfere with the expression of the target gene to a certain extent, and among them, it was found that the #3 siRNA of siPRDX5, and the #3 and #4 siRNAs of siGPX4 have more excellent ability to interfere with gene expression.

[0044] Example 3 Modification of siRNA and verification of interference ability The #3 siRNA of siPRDX5, the #3 siRNA of siGPX4, and the #4 siRNA of siGPX4 screened in Example 2 were subjected to base modification and synthesized by a biological company.

[0045] Here, A-, U-, C-, and G- respectively represent phosphorothioate-modified ribonucleotides A, U, C, and G, (A), (U), (C), and (G) respectively represent 2'-F-modified ribonucleotides A, U, C, and G, [A], [U], [C], and [G] respectively represent 2'-OMe-modified ribonucleotides A, U, C, and G, (A) , (U) , (C) , and (G) respectively represent 2'-Ara-F-modified ribonucleotides A, U, C, and G, [A] , [U] , [C] , and [G] respectively represent 2'-O-MOE-modified ribonucleotides A, U, C, and G, A*, U*, C*, and G* respectively represent m 6 A-modified ribonucleotides A, U, C, and G, A', U', C', and G' respectively represent m 5 C-modified ribonucleotides A, U, C, and G.

[0046] siPRDX5#3siRNA was similar to that of siPRDX5-1~siPRDX5-16.

[0047] siPRDX5-1 5' (C)-[A]-(G)[A](C)[U](U)[A](UUA)[C](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C)[A](U)[C](U)[A](G)[UAA](U)[A](A)[G](U)[C](U)[G]-[U]-[C] 3'; siPRDX5-2 5′ (C)–[ A ]–(G)[ A ](C)[ U ](U)[ A ](UUA)[ C ](U)[ A ](G)[ A ](U)[ G ](A)[ U ](T) 3′; 5' [ A ]–(A)–[ U ](C)[ A ](U)[ C ](U)[ A ](G)[ UAA ](U)[ A ](A)[ G ](U)[ C ](U)[ G ]–[ U ]–[ C ] 3'; siPRDX5-3 5' ( C )-[A]-( G )[A]( C )[U]( U )[A]( UUA )[C]( U )[A]( G )[A]( U )[G]( A )[U]( T ) 3'; 5' [A]-( A )-[U]( C )[A]( U)[C]( U )[A]( G )[UAA]( U )[A]( A )[G]( U )[C]( U )[G]-[U]-[C] 3'; siPRDX5-4 5' ( C )-[ A ]-( G )[ A ]( C )[ U ]( U )[ A ]( UUA )[ C ]( U )[ A ]( G )[ A ]( U )[ G ]( A )[ U ]( T ) 3'; 5' [ A ]-( A )-[ U ]( C )[ A ]( U )[ C ]( U )[ A ]( G )[ UAA ]( U )[ A ]( A )[ G ]( U )[ C ]( U )[ G ]-[ U ]-[ C ] 3'; siPRDX5-5 5' (C)-[A*]-(G)[A*](C)[U](U)[A*](UUA*)[C](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5' [A*]-(A*)-[U](C)[A*](U)[C](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C](U)[G]-[U]-[C] 3'; siPRDX5-6 5' (C')-[A]-(G)[A](C')[U](U)[A](UUA)[C'](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C')[A](U)[C'](U)[A](G)[UAA](U)[A](A)[G](U)[C'](U)[G]-[U]-[C'] 3'; siPRDX5-7 5' (C)-[ A *]-(G)[ A *](C)[ U ](U)[ A *](UUA*)[ C ](U)[ A *](G)[ A *](U)[ G ](A*)[ U ](T) 3'; 5'[ A *]-(A*)-[ U ](C)[ A *](U)[ C ](U)[ A *](G)[ UA * A *](U)[ A *](A*)[ G ](U)[ C ](U)[ G ]-[ U ]-[ C ] 3'; siPRDX5-8 5' ( C )-[A*]-( G )[A*]( C )[U]( U )[A*]( UUA *)[C]( U )[A*]( G )[A*]( U )[G]( A *)[U]( T ) 3'; 5' [A*]-( A *)-[U]( C )[A*]( U )[C]( U )[A*](G )[UA*A*]( U )[A*]( A *)[G]( U )[C]( U )[G]-[U]-[C] 3'; siPRDX5-9 5' ( C )-[ A *]-( G )[ A *]( C )[ U ]( U )[ A *]( UUA *)[ C ]( U )[ A *]( G )[ A *]( U )[ G ]( A *)[ U ]( T ) 3'; 5' [ A *]-( A *)-[ U ]( C )[ A *]( U )[ C ]( U )[ A *]( G )[ UA * A *]( U )[ A *]( A *)[ G ]( U )[ C ]( U )[ G ]-[ U ]-[ C ] 3'; siPRDX5-10 5' (C')-[ A ]-(G)[ A ](C')[ U ](U)[ A ](UUA)[ C '](U)[ A ](G)[ A ](U)[ G ](A)[U ](T) 3'; 5' [ A ]-(A)-[ U ](C')[ A ](U)[ C '](U)[ A ](G)[ UAA ](U)[ A ](A)[ G ](U)[ C '](U)[ G ]-[ U ]-[ C '] 3'; siPRDX5-11 5' ( C ')-[A]-( G )[A]( C ')[U]( U )[A]( UUA )[C']( U )[A]( G )[A]( U )[G]( A )[U]( T ) 3'; 5' [A]-( A )-[U]( C ')[A]( U )[C']( U )[A]( G )[UAA]( U )[A]( A )[G]( U )[C']( U )[G]-[U]-[C'] 3'; siPRDX5-12 5' ( C ')-[ A ]-( G )[ A ]( C ')[ U ]( U )[ A ]( UUA )[ C ']( U )[ A ]( G )[ A ]( U )[ G ]( A )[ U ](T ) 3'; 5' [ A ]-( A )-[ U ]( C ')[ A ]( U )[ C ']( U )[ A ]( G )[ UAA ]( U )[ A ]( A )[ G ]( U )[ C ']( U )[ G ]-[ U ]-[ C '] 3'; siPRDX5-13 5' (C')-[A*]-(G)[A*](C')[U](U)[A*](UUA*)[C'](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5'[A*]-(A*)-[U](C')[A*](U)[C'](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C'](U)[G]-[U]-[C']3'; siPRDX5-14 5' (C')-[ A *]-(G)[ A *](C')[ U ](U)[ A *](UUA*)[ C '](U)[ A *](G)[ A *](U)[ G ](A*)[ U ](T) 3'; 5'[ A *]-(A*)-[ U ](C')[ A *](U)[ C '](U)[ A *](G)[ UA * A *](U)[ A *](A*)[ G ](U)[ C '](U)[G ]-[ U ]-[ C ']3'; siPRDX5-15 5' ( C ')-[A*]-( G )[A*]( C ')[U]( U )[A*]( UUA *)[C']( U )[A*]( G )[A*]( U )[G]( A *)[U]( T ) 3'; 5'[A*]-( A *)-[U]( C ')[A*]( U )[C']( U )[A*]( G )[UA*A*]( U )[A*]( A *)[G]( U )[C']( U )[G]-[U]-[C']3'; siPRDX5-16 5' ( C ')-[ A *]-( G )[ A *]( C ')[ U ]( U )[ A *]( UUA *)[ C ']( U )[ A *]( G )[ A *]( U )[ G ]( A *)[ U ]( T ) 3'; 5'[ A *]-( A *)-[ U ]( C ')[ A *]( U )[ C ']( U )[ A *]( G )[UA * A *]( U )[ A *]( A *)[ G ]( U )[ C ']( U )[ G ]-[ U ]-[ C ']3'; The sequences of siGPX4 with base modification of #3siRNA are shown below in siGPX4-1 to siGPX4-16.

[0048] siGPX4-1 5' (A)-[C]-(A)[A](C)[G](U)[C](AAA)[U](U)[C](G)[A](U)[A](U)[G](U) 3'; 5' [A]-(C)-[A](U)[A](U)[C](G)[A](A)[UUU](G)[A](C)[G](U)[U](G)[U]-[A]-[C] 3'; siGPX4-2 5' (A)-[ C ]-(A)[ A ](C)[ G ](U)[ C ](AAA)[ U ](U)[ C ](G)[ A ](U)[ A ](U)[ G ](U) 3'; 5' [ A ]-(C)-[ A ](U)[ A ](U)[ C ](G)[ A ](A)[ UUU ](G)[ A ](C)[ G ](U)[ U ](G)[ U ]-[ A ]-[ C ] 3'; siGPX4-3 5' ( A )-[C]-( A )[A]( C)[G]( U )[C]( AAA )[U]( U )[C]( G )[A]( U )[A]( U )[G]( U ) 3’; 5’ [A]-( C )-[A]( U )[A]( U )[C]( G )[A]( A )[UUU]( G )[A]( C )[G]( U )[U]( G )[U]-[A]-[C] 3’; siGPX4-4 5’ ( A )-[ C ]-( A )[ A ]( C )[ G ]( U )[ C ]( AAA )[ U ]( U )[ C ]( G )[ A ]( U )[ A ]( U )[ G ]( U ) 3’; 5’ [ A ]-( C )-[ A ]( U )[ A ]( U )[ C ]( G )[ A ]( A )[ UUU ]( G )[ A ]( C )[ G ]( U )[ U ]( G )[ U ]-[ A ]-[ C ] 3’; siGPX4-5 5’ (A*)-[C]-(A*)[A*](C)[G](U)[C](A*A*A*)[U](U)[C](G)[A*](U)[A*](U)[G](U) 3’; 5’ [A*]-(C)-[A*](U)[A*](U)[C](G)[A*](A*)[UUU](G)[A*](C)[G](U)[U](G)[U]-[A*]-[C] 3’; siGPX4-6 5’ (A)-[C’]-(A)[A](C’)[G](U)[C’](AAA)[U](U)[C’](G)[A](U)[A](U)[G](U) 3’; 5’ [A]-(C’)-[A](U)[A](U)[C’](G)[A](A)[UUU](G)[A](C’)[G](U)[U](G)[U]-[A]-[C’] 3’; siGPX4-7 5’ (A*)-[ C ]-(A*)[ A *](C)[ G ](U)[ C ](A*A*A*)[ U ](U)[ C ](G)[ A *](U)[ A *](U)[ G ](U) 3’; 5’ [ A *]-(C)-[ A *](U)[ A *](U)[ C ](G)[ A *](A*)[ UUU ](G)[ A *](C)[ G ](U)[ U ](G)[ U ]-[ A *]-[ C ] 3’; siGPX4-8 5’ ( A *)-[C]-( A *)[A*]( C )[G]( U )[C]( A * A* A *)[U]( U )[C]( G )[A*]( U )[A*]( U )[G]( U ) 3’; 5’ [A*]-( C )-[A*]( U )[A*]( U )[C]( G )[A*]( A *)[UUU]( G )[A*]( C )[G]( U )[U]( G )[U]-[A*]-[C] 3’; siGPX4-9 5’ ( A *)-[ C ]-( A *)[ A *]( C )[ G ]( U )[ C ]( A * A * A *)[ U ]( U )[ C ]( G )[ A *]( U )[ A *]( U )[ G ]( U ) 3’; 5’ [ A *]-( C )-[ A *]( U )[ A *]( U )[ C ]( G )[ A *]( A *)[ UUU ]( G )[ A *]( C )[ G ]( U )[ U ]( G )[ U ]-[ A*]-[ C ] 3’; siGPX4-10 5’ (A)-[ C ’]-(A)[ A ](C’)[ G ](U)[ C ’](AAA)[ U ](U)[ C ’](G)[ A ](U)[ A ](U)[ G ](U) 3’; 5’ [ A ]-(C’)-[ A ](U)[ A ](U)[ C ’](G)[ A ](A)[ UUU ](G)[ A ](C’)[ G ](U)[ U ](G)[ U ]-[ A ]-[ C ’] 3’; siGPX4-11 5’ ( A )-[C’]-( A )[A]( C ’)[G]( U )[C’]( AAA )[U]( U )[C’]( G )[A]( U )[A]( U )[G]( U ) 3’; 5’ [A]-( C ’)-[A]( U )[A]( U )[C’]( G )[A]( A )[UUU]( G )[A]( C ’)[G]( U )[U]( G )[U]-[A]-[C’] 3’; siGPX4-12 5’ ( A )-[ C ’]-( A )[ A ](C ’)[ G ]( U )[ C ’]( AAA )[ U ]( U )[ C ’]( G )[ A ]( U )[ A ]( U )[ G ]( U ) 3’; 5’ [ A ]-( C ’)-[ A ]( U )[ A ]( U )[ C ’]( G )[ A ]( A )[ UUU ]( G )[ A ]( C ’)[ G ]( U )[ U ]( G )[ U ]-[ A ]-[ C ’] 3’; siGPX4-13 5’ (A*)-[C’]-(A*)[A*](C’)[G](U)[C’](A*A*A*)[U](U)[C’](G)[A*](U)[A*](U)[G](U) 3’; 5’ [A*]-(C’)-[A*](U)[A*](U)[C’](G)[A*](A*)[UUU](G)[A*](C’)[G](U)[U](G)[U]-[A*]-[C’] 3’; siGPX4-14 5’ (A*)-[ C ’]-(A*)[ A *](C’)[ G ](U)[ C ’](A*A*A*)[ U ](U)[ C ’](G)[ A *](U)[ A *](U)[ G](U) 3’; 5’ [ A *]-(C’)-[ A *](U)[ A *](U)[ C ’](G)[ A *](A*)[ UUU ](G)[ A *](C’)[ G ](U)[ U ](G)[ U ]-[ A *]-[ C ’] 3’; siGPX4-15 5’ ( A *)-[C’]-( A *)[A*]( C ’)[G]( U )[C’]( A * A * A *)[U]( U )[C’]( G )[A*]( U )[A*]( U )[G]( U ) 3’; 5’ [A*]-( C ’)-[A*]( U )[A*]( U )[C’]( G )[A*]( A *)[UUU]( G )[A*]( C ’)[G]( U )[U]( G )[U]-[A*]-[C’] 3’; siGPX4-16 5’ ( A *)-[ C ’]-( A *)[ A *]( C ’)[ G ]( U )[ C ’]( A * A * A *)[ U ]( U )[ C ’]( G )[ A*]( U )[ A *]( U )[ G ]( U ) 3'; 5' [ A *]-( C ')-[ A *]( U )[ A *]( U )[ C ']( G )[ A *]( A *)[ UUU ]( G )[ A *]( C ')[ G ]( U )[ U ]( G )[ U ]-[ A *]-[ C '] 3'; The sequences of siGPX4 with base modification of #4siRNA are shown in siGPX4-(1) to siGPX4-(16).

[0049] siGPX4-(1) 5' (G)-[U]-(G)[A](G)[G](C)[A](AGA)[C](C)[G](A)[A](G)[U](A)[A](A) 3'; 5' [U]-(U)-[U](A)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A)[C]-[U]-[G] 3'; siGPX4-(2) 5' (G)-[ U ]-(G)[ A ](G)[ G ](C)[ A (AGA) C ](C)[ G ](A)[ A ](G)[ U ](A)[ A ](A) 3'; 5' [ U ]-(U)-[ U ](A)[ C](U)[ U ](C)[ G ](G)[ UCU ](U)[ G ](C)[ C ](U)[ C ](A)[ C ]-[ U ]-[ G ] 3’; siGPX4-(3) 5’ ( G )-[U]-( G )[A]( G )[G]( C )[A]( AGA )[C]( C )[G]( A )[A]( G )[U]( A )[A]( A ) 3’; 5’ [U]-( U )-[U]( A )[C]( U )[U]( C )[G]( G )[UCU]( U )[G]( C )[C]( U )[C]( A )[C]-[U]-[G] 3’; siGPX4-(4) 5’ ( G )-[ U ]-( G )[ A ]( G )[ G ]( C )[ A ]( AGA )[ C ]( C )[ G ]( A )[ A ]( G )[ U ]( A )[ A ]( A ) 3’; 5’ [ U ]-( U )-[ U ]( A )[C ]( U )[ U ]( C )[ G ]( G )[ UCU ]( U )[ G ]( C )[ C ]( U )[ C ]( A )[ C ]-[ U ]-[ G ] 3’; siGPX4-(5) 5’ (G)-[U]-(G)[A*](G)[G](C)[A*](A*GA*)[C](C)[G](A*)[A*](G)[U](A*)[A*](A*) 3’; 5’ [U]-(U)-[U](A*)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A*)[C]-[U]-[G] 3’; siGPX4-(6) 5’ (G)-[U]-(G)[A](G)[G](C’)[A](AGA)[C’](C’)[G](A)[A](G)[U](A)[A](A) 3’; 5’ [U]-(U)-[U](A)[C’](U)[U](C’)[G](G)[UC’U](U)[G](C’)[C’](U)[C’](A)[C’]-[U]-[G] 3’; siGPX4-(7) 5’ (G)-[ U ]-(G)[ A *](G)[ G ](C)[ A *](A*GA*)[ C ](C)[ G ](A*)[ A *](G)[ U ](A*)[ A *](A*) 3’; 5’ [ U ]-(U)-[ U ](A*)[ C ](U)[ U ](C)[ G ](G)[UCU ](U)[ G ](C)[ C ](U)[ C ](A*)[ C ]-[ U ]-[ G ] 3’; siGPX4-(8) 5’ ( G )-[U]-( G )[A*]( G )[G]( C )[A*]( A * GA *)[C]( C )[G]( A *)[A*]( G )[U]( A *)[A*]( A *) 3’; 5’ [U]-( U )-[U]( A *)[C]( U )[U]( C )[G]( G )[UCU]( U )[G]( C )[C]( U )[C]( A *)[C]-[U]-[G] 3’; siGPX4-(9) 5’ ( G )-[ U ]-( G )[ A *]( G )[ G ]( C )[ A *]( A * GA *)[ C ]( C )[ G ]( A *)[ A *]( G )[ U ]( A *)[ A *]( A *) 3’; 5’ [ U ]-( U )-[ U ]( A *)[C ]( U )[ U ]( C )[ G ]( G )[ UCU ]( U )[ G ]( C )[ C ]( U )[ C ]( A *)[ C ]-[ U ]-[ G ] 3’; siGPX4-(10) 5’ (G)-[ U ]-(G)[ A ](G)[ G ](C’)[ A ](AGA)[ C ’](C’)[ G ](A)[ A ](G)[ U ](A)[ A ](A) 3’; 5’ [ U ]-(U)-[ U ](A)[ C ’](U)[ U ](C’)[ G ](G)[ UC ’ U ](U)[ G ](C’)[ C ’](U)[ C ’](A)[ C ’]-[ U ]-[ G ] 3’; siGPX4-(11) 5’ ( G )-[U]-( G )[A]( G )[G]( C ’)[A]( AGA )[C’]( C ’)[G]( A )[A]( G )[U]( A )[A]( A ) 3’; 5’ [U]-( U )-[U](A )[C’]( U )[U]( C ’)[G]( G )[UC’U]( U )[G]( C ’)[C’]( U )[C’]( A )[C’]-[U]-[G] 3’; siGPX4-(12) 5’ ( G )-[ U ]-( G )[ A ]( G )[ G ]( C ’)[ A ]( AGA )[ C ’]( C ’)[ G ]( A )[ A ]( G )[ U ]( A )[ A ]( A ) 3’; 5’ [ U ]-( U )-[ U ]( A )[ C ’]( U )[ U ]( C ’)[ G ]( G )[ UC ’ U ]( U )[ G ]( C ’)[ C ’]( U )[ C ’]( A )[ C ’]-[ U ]-[ G ] 3’; siGPX4-(13) 5’ (G)-[U]-(G)[A*](G)[G](C’)[A*](A*GA*)[C’](C’)[G](A*)[A*](G)[U](A*)[A*](A*) 3’; 5’ [U]-(U)-[U](A*)[C’](U)[U](C’)[G](G)[UC’U](U)[G](C’)[C’](U)[C’](A*)[C’]-[U]-[G] 3’; siGPX4-(14) 5’ (G)-[ U ]-(G)[ A *](G)[ G ](C’)[ A *](A*GA*)[ C ’](C’)[ G ](A*)[ A *](G)[ U ](A*)[ A *](A*) 3’; 5’ [ U ]-(U)-[ U ](A*)[ C ’](U)[ U ](C’)[ G ](G)[ UC ’ U ](U)[ G ](C’)[ C ’](U)[ C ’](A*)[ C ’]-[ U ]-[ G ] 3’; siGPX4-(15) 5’ ( G )-[U]-( G )[A*]( G )[G]( C ’)[A*]( A * GA *)[C’]( C ’)[G]( A *)[A*]( G )[U]( A *)[A*]( A *) 3’; 5’ [U]-( U )-[U]( A *)[C’]( U )[U]( C ’)[G]( G )[UC’U]( U )[G]( C ’)[C’]( U )[C’]( A*)[C’]-[U]-[G] 3’; siGPX4-(16) 5’ ( G )-[ U ]-( G )[ A *]( G )[ G ]( C ’)[ A *]( A * GA *)[ C ’]( C ’)[ G ]( A *)[ A *]( G )[ U ]( A *)[ A *]( A *) 3’; 5’ [ U ]-( U )-[ U ]( A *)[ C ’]( U )[ U ]( C ’)[ G ]( G )[ UC ’ U ]( U )[ G ]( C ’)[ C ’]( U )[ C ’]( A *)[ C ’]-[ U ]-[ G ] 3’。

[0050] As shown in Figures 2 and 3, and Tables 1 and 2, LNCaP cells and MyC-CaP cells, as well as LNCaP-DTP and MyC-CaP-DTP cells derived therefrom, were treated with the modified siRNAs using the same method as in Example 2. The results summarized in Tables 1 and 2 show that the sequences siPRDX5-1 to siPRDX5-16 (with #3 siRNA modified from siPRDX5), siGPX4-1 to 1-siGPX4-16 (with #3 siRNA modified from siGPX4), and #4siGPX4-(1) to #4siGPX4-(16) (with #4 siRNA modified from siGPX4) exhibited strong interference activity against both drug-resistant and non-drug-resistant LNCaP and MyC-CaP cells. Furthermore, #3siGPX4 showed higher interference efficiency than #4siGPX4. It was also found that the interference efficiency was higher against human cells than against mouse cells.

[0051] [Table 1]

[0052] [Table 2] Note: "NO." indicates the sequence number of the base modification.

[0053] Example 4: Killing ability of siPRDX5 and siGPX4 against reversible drug-resistant prostate cancer cells Furthermore, when the modified #3siPRDX5-1~16, #3siPRDX4-1~16, or #3siPRDX5-1-16+#3siGPX4-1-16 synthesized in Example 3 were transmitted using the CCK8 method to drug-resistant LNCaP-DTP and MyC-CaP-DTP cells, as well as ENZ-resistant 22Rv1 cells, it was shown that the nucleic acid drugs have in vitro antitumor activity in drug-resistant cells.

[0054] 1. Experimental Method Drug-resistant cells LNCaP-DTP, MyC-CaP-DTP, and ENZ-resistant 22Rv1 cells were seeded in a 96-well plate containing fresh 1640 medium. After adhesion, 1-16 siPRDX5, siGPX4, siPRDX5+siGPX4, and negative control si-neg were added. If interference was successful, the transfection mixture was aspirated and replaced with standard complete medium (1640 medium containing 10% fetal bovine serum and 1% biantibody). The cells were incubated in an incubator for 24 hours and finally detected by CCK8. 450 The cell viability rate was calculated based on the values.

[0055] 2. The results are shown in Figure 5. Figure 5A is a bar graph of the relative survival rate of human prostate cancer drug-resistant cells LNCaP-DTP administered with 1 to 16 modified siRNAs of siPRDX5, Figure 5B is a bar graph of the relative survival rate of mouse prostate cancer drug-resistant cells MyC-CaP-DTP administered with siRNAs of siPRDX5-1 to 16, Figure 5C is a bar graph of the relative survival rate of human prostate cancer ENZ drug-resistant cells 22Rv1 administered with siRNAs of siPRDX5-1 to 16, Figure 5D is a bar graph of the relative survival rate of human prostate cancer drug-resistant cells LNCaP-DTP administered with siRNAs of siGPX4-1 to 16, Figure 5E is a bar graph of the relative survival rate of mouse prostate cancer drug-resistant cells MyC-CaP-DTP administered with siRNAs of siGPX4-1 to 16, Figure 5F is a bar graph of siGP Figure 5G is a bar graph showing the relative survival rate of human prostate cancer ENZ drug-resistant cells 22Rv1 administered with each of the siRNAs X4-1 to X4-16. Figure 5H is a bar graph showing the relative survival rate of human prostate cancer drug-resistant cells LNCaP-DTP administered with each of the siRNAs siGPX4-1 to X4-16 and siPRDX5-1 to X4-16. Figure 5I is a bar graph showing the relative survival rate of mouse prostate cancer drug-resistant cells MyC-CaP-DTP administered with each of the siRNAs siGPX4-1 to X4-16 and siPRDX5-1 to X4-16.

[0056] As a result, in drug-resistant DTP cells obtained by administering ENZ for 9 consecutive days, there was no significant inhibitory effect when ENZ was added again. However, when the nucleic acid drugs siPRDX5 or siGPX4, which have different sequences, were added, the inhibition rate was approximately 50% in all three cell types. Furthermore, when siPRDX5 + siGPX4 was added, a stronger inhibitory effect on the growth of the three cell types was observed compared to the addition of one siRNA alone. This indicates that siPRDX5 and siGPX4 have a clear killing effect on drug-resistant prostate cancer cells and the ability to inhibit the growth of drug-resistant cells, and that a stronger effect can be obtained by using these two siRNAs in combination.

[0057] Example 5: Effect of combination therapy with ENZ and nucleic acid drugs on a mouse model of C-MYC overexpressing prostate cancer in which resistance to CRPC was induced by continuous administration of ENZ. Furthermore, we further described the effects of combining ENZ with modified #3siPRDX5 and #3siGPX4 in mice that relapsed after chemical castration (i.e., continued administration of ENZ) in a mouse model of prostate cancer.

[0058] 1. Experimental Method We constructed a mouse model of spontaneously occurring prostate cancer that overexpresses C-MYC (Hi-Myc). The mice progressed to mPIN / cancer transition at 4 months of age. At this point, the mice were randomly divided into an NC control group (intragastric administration) and an ENZ administration group. The ENZ administration group received 10 mg / kg of ENZ intragastricly every 3 days for a total of 30 days. Subsequently, several mice were killed by neck dislocation, and their prostate cancers were harvested, photographed, and weighed. The results showed that ENZ significantly reduced symptoms, and the prostate weight was reduced by half compared to the NC control group. Next, administration was continued to the remaining mice for 60 days according to the same method. Recurrence was observed in the ENZ group, and we successfully obtained CRPC mice. Subsequently (i.e., at 7 months of age), mice were randomly divided into the following groups: NC control group (always administered the solvent intragastricly), ENZ monotherapy group, siPRDX5-1 nucleic acid drug group, ENZ and siPRDX5-1 combination group, siPRDX5-16 nucleic acid drug group, ENZ and siPRDX5-16 combination group, siGPX4-1 nucleic acid drug group, ENZ and siGPX4-1 combination group, siGPX4-16 nucleic acid drug group, ENZ and siGPX4-16 combination group, siPRDX5-16 + siGPX4-16 nucleic acid drug group, and ENZ and siPRDX5-16 + siGPX4-16 combination group, and the corresponding treatment was administered. However, NC and ENZ continued to be administered intragastricly, while siPRDX5-1, siPRDX5-16, and siGPX4-1 and siGPX4-16 were all administered by tail vein injection every three days, maintaining ENZ at 10 mg / kg and siPRDX5-1 / 16 and siGPX4-1 / 16 at 100 nM each time, for a total of 30 days. Subsequently, the mice were killed by neck dislocation, their prostate cancers were collected, photographed, weighed, and subjected to experiments such as HE staining.

[0059] 2. The results are shown in Figures 6 and 7. In Figure 6, Figure 6A shows the change in body weight of mice in each group as administration progresses, Figure 6B is a comparative view of photographs taken after prostatectomy of mice in each group, and Figure 6C shows the change in prostate weight of mice in each group as administration progresses, with * indicating whether the decrease in prostate weight in the other ENZ groups and the siPRDX5 and siGPX4 administration groups was significant compared to the NC group. In Figure 7, Figure 7A is an HE stained view of prostate tissue sections of mice in each group, and Figure 7B is a bar graph showing the relative change in PRDX5 enzyme activity detected in the prostate tissue of mice in each group, with * indicating whether the decrease in PRDX5 enzyme activity in the other siPRDX5 and siGPX4 administration groups was significant compared to the ENZ group.

[0060] Table 3 shows the results of using the drug alone and in combination.

[0061] [Table 3]

[0062] Combining Figure 6 and Table 3, it was found that the average prostate weight of drug-resistant CRPC mice administered ENZ intragastricly for 30 consecutive days was 66.67 mg, while the average prostate weight of the NC control group was 115.6 mg. At this point, the prostate weight of the siPRDX5-1 group decreased to 53.99 mg after 30 days of administration, and the prostate weight of the siPRDX5-1 and ENZ administration group decreased to 49.95 mg, demonstrating that the siPRDX5 nucleic acid drug has a therapeutic effect in inhibiting tumor growth in CRPC mice. The prostate weight of the siGPX4-1 group after 30 days of administration was 57.81 mg, and the prostate weight of the ENZ and siGPX4-1 combination group was 49.40 mg, demonstrating that the siGPX4 nucleic acid drug also has a therapeutic effect in inhibiting tumor growth in CRPC mice. After 30 days of administration, the prostate weight decreased to 47.28 mg in the siPRDX5-16 + siGPX4-16 group, while the prostate weight decreased to 39.35 mg in the siPRDX5-16 + siGPX4-16 + ENZ combination group, demonstrating a more pronounced efficacy of the siPRDX5 + siGPX4 nucleic acid drug.

[0063] The combination of ENZ with siPRDX5 or siGPX4 showed superior efficacy compared to siPRDX5 or siGPX4 alone, reducing prostate weight to approximately 50 mg. Furthermore, siPRDX5 or siGPX4 alone was found to be superior to ENZ alone after drug resistance. siPRDX5 or siGPX4 alone demonstrated inhibitory effects against drug-resistant CRPC mice. The combination of ENZ with two nucleic acid drugs showed the highest efficacy, reducing prostate weight to approximately 40 mg.

[0064] Method for detecting PRDX5 enzyme activity The PRDX5 enzyme activity was detected using a thioredoxin system. Specifically, the rate of H2O2 degradation was measured by detecting the decrease in A340 due to the oxidation of NADPH. The experiment was performed using 150 μL of a reaction mixture containing 50 mM EPES-NaOH (pH=7), 200 μM NADPH, 760 nM mouse TXNRD1, 11 μM human TRX, and PRDX5 at various concentration gradients. The mixture was incubated at 37°C for 5 minutes, and the reaction was initiated by adding 500 μM H2O2.

[0065] Table 4 shows the results of PRDX5 enzyme activity in the prostate gland from various groups.

[0066] [Table 4]

[0067] HE staining results of tissue sections (Figure 7) showed that, with concomitant administration, prostate tumors in CRPC exhibited clear death, regression, and fibrosis. Combining the detection results of PRDX5 enzyme activity in the prostate of various groups (Table 4), it was found that with continuous intragastric administration of ENZ, when mice developed CRPC, PRDX5 enzyme activity increased significantly, reaching a relative value of 4.56. On the other hand, with the use of the nucleic acid drug siPRDX5-1, enzyme activity was significantly inhibited, reaching a relative value of 2.025, and especially with concomitant use, it reached a relative value of 1.465. With the use of the nucleic acid drug siGPX4-1, PRDX5 enzyme activity was also significantly inhibited, reaching a relative value of 1.810, and with concomitant use, it reached a relative value of 1.600. These two nucleic acid drugs were proven to have a clear interference effect when used individually. When the two nucleic acid drugs were administered simultaneously, the relative enzyme activity of PRDX5 in the siPRDX5-16 + siGPX4-16 group reached 1.350. When administered in combination with ENZ, the enzyme activity was the lowest at 0.955. This indicates that simultaneous administration of the siPRDX5-16 + siGPX4-16 nucleic acid drugs can achieve the best inhibition of PRDX5 enzyme activity and the best inhibition of prostate tumor formation in CRPC mice.

[0068] Although the present invention has been disclosed above by preferred embodiments, these are not intended to limit the invention, and those skilled in the art can make various changes and modifications without departing from the spirit and scope of the invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A double-stranded siRNA molecule, wherein the double-stranded siRNA molecule is (A) A siRNA molecule that inhibits PRDX5 gene expression, and which has a double-stranded siRNA molecule in which the single-stranded RNA shown in SEQ ID NO. 1 and the single-stranded RNA shown in SEQ ID NO. 2 are complementary. A double-stranded siRNA molecule characterized by the following features.

2. A composition, (A) A siRNA molecule that inhibits PRDX5 gene expression, comprising a double-stranded siRNA molecule in which the single-stranded RNA shown in SEQ ID NO. 1 and the single-stranded RNA shown in SEQ ID NO. 2 are complementary. (B) A siRNA molecule that inhibits GPX4 gene expression, having a double-stranded siRNA molecule in which the RNA single strand shown in SEQ ID NO. 3 and the RNA single strand shown in SEQ ID NO. 4 are complementary, or having a double-stranded siRNA molecule in which the RNA single strand shown in SEQ ID NO. 5 and the RNA single strand shown in SEQ ID NO. 6 are complementary. A composition characterized by containing [a certain substance].

3. The double-stranded siRNA molecule according to claim 1, characterized in that at least one nucleotide in the double-stranded siRNA molecule is a modified nucleotide.

4. The double-stranded siRNA molecule according to claim 1, characterized in that all nucleotides in the double-stranded siRNA molecule are modified nucleotides.

5. The aforementioned modifications are phosphorothioate, 2'-F, 2'-OMe, 2'-Ara-F, 2'-O-MOE, m ６ A or m ５ The double-stranded siRNA molecule according to claim 3, characterized in that it is at least one selected from C.

6. The composition according to claim 2, characterized in that the double-stranded siRNA molecule is any of (A1) to (C16) below. (A1) 5' (C) - [A] - (G) [A] (C) [U] (U) [A] (UUA) [C] (U) [A] (G) [A] (U) [G] (A) [U] (T) 3'; 5' [A] - (A) - [U] (C) [A] (U) [C] (U) [A] (G) [UAA] (U) [A] (A) [G] (U) [C] (U) [G] - [U] - [C] 3'; (A2) 5' (C) - [A] - (G) [A] (C) [U] (U) [A] (UUA) [C] (U) [A] (G) [A] (U) [G] (A) [U] (T) 3'; 5' [A] - (A) - [U] (C) [A] (U) [C] (U) [A] (G) [UAA] (U) [A] (A) [G] (U) [C] (U) [G] - [U] - [C] 3'; (A3) 5' (C)-[A]-(G)[A](C)[U](U)[A](UUA)[C](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C)[A](U)[C](U)[A](G)[UAA](U)[A](A)[G](U)[C](U)[G]-[U]-[C] 3'; (A4) 5' (C)-[A]-(G)[A](C)[U](U)[A](UUA)[C](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C)[A](U)[C](U)[A](G)[UAA](U)[A](A)[G](U)[C](U)[G]-[U]-[C] 3'; (A5) 5' (C)-[A*]-(G)[A*](C)[U](U)[A*](UUA*)[C](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5' [A*]-(A*)-[U](C)[A*](U)[C](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C](U)[G]-[U]-[C] 3'; (A6) 5' (C')-[A]-(G)[A](C')[U](U)[A](UUA)[C'](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C')[A](U)[C'](U)[A](G)[UAA](U)[A](A)[G](U)[C'](U)[G]-[U]-[C'] 3'; (A7) 5' (C)-[A*]-(G)[A*](C)[U](U)[A*](UUA*)[C](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5'[A*]-(A*)-[U](C)[A*](U)[C](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C](U)[G]-[U]-[C] 3'; (A8) 5' (C)-[A*]-(G)[A*](C)[U](U)[A*](UUA*)[C](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5' [A*]-(A*)-[U](C)[A*](U)[C](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C](U)[G]-[U]-[C] 3'; (A9) 5' (C)-[A*]-(G)[A*](C)[U](U)[A*](UUA*)[C](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5' [A*]-(A*)-[U](C)[A*](U)[C](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C](U)[G]-[U]-[C] 3'; (A10) 5' (C')-[A]-(G)[A](C')[U](U)[A](UUA)[C'](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C')[A](U)[C'](U)[A](G)[UAA](U)[A](A)[G](U)[C'](U)[G]-[U]-[C'] 3'; (A11) 5' (C')-[A]-(G)[A](C')[U](U)[A](UUA)[C'](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C')[A](U)[C'](U)[A](G)[UAA](U)[A](A)[G](U)[C'](U)[G]-[U]-[C'] 3'; (A12) 5' (C')-[A]-(G)[A](C')[U](U)[A](UUA)[C'](U)[A](G)[A](U)[G](A)[U](T) 3'; 5' [A]-(A)-[U](C')[A](U)[C'](U)[A](G)[UAA](U)[A](A)[G](U)[C'](U)[G]-[U]-[C'] 3'; (A13) 5' (C')-[A*]-(G)[A*](C')[U](U)[A*](UUA*)[C'](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5'[A*]-(A*)-[U](C')[A*](U)[C'](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C'](U)[G]-[U]-[C']3'; (A14) 5' (C')-[A*]-(G)[A*](C')[U](U)[A*](UUA*)[C'](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5'[A*]-(A*)-[U](C')[A*](U)[C'](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C'](U)[G]-[U]-[C']3'; (A15) 5' (C')-[A*]-(G)[A*](C')[U](U)[A*](UUA*)[C'](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5'[A*]-(A*)-[U](C')[A*](U)[C'](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C'](U)[G]-[U]-[C']3'; (A16) 5' (C')-[A*]-(G)[A*](C')[U](U)[A*](UUA*)[C'](U)[A*](G)[A*](U)[G](A*)[U](T) 3'; 5'[A*]-(A*)-[U](C')[A*](U)[C'](U)[A*](G)[UA*A*](U)[A*](A*)[G](U)[C'](U)[G]-[U]-[C']3'; (B1) 5' (A)-[C]-(A)[A](C)[G](U)[C](AAA)[U](U)[C](G)[A](U)[A](U)[G](U) 3'; 5' [A]-(C)-[A](U)[A](U)[C](G)[A](A)[UUU](G)[A](C)[G](U)[U](G)[U]-[A]-[C] 3'; (B2) 5' (A)-[C]-(A)[A](C)[G](U)[C](AAA)[U](U)[C](G)[A](U)[A](U)[G](U) 3'; 5' [A]-(C)-[A](U)[A](U)[C](G)[A](A)[UUU](G)[A](C)[G](U)[U](G)[U]-[A]-[C] 3'; (B3) 5' (A)-[C]-(A)[A](C)[G](U)[C](AAA)[U](U)[C](G)[A](U)[A](U)[G](U) 3'; 5' [A]-(C)-[A](U)[A](U)[C](G)[A](A)[UUU](G)[A](C)[G](U)[U](G)[U]-[A]-[C] 3'; (B4) 5' (A)-[C]-(A)[A](C)[G](U)[C](AAA)[U](U)[C](G)[A](U)[A](U)[G](U) 3'; 5' [A]-(C)-[A](U)[A](U)[C](G)[A](A)[UUU](G)[A](C)[G](U)[U](G)[U]-[A]-[C] 3'; (B5) 5' (A*)-[C]-(A*)[A*](C)[G](U)[C](A*A*A*)[U](U)[C](G)[A*](U)[A*](U)[G](U) 3'; 5' [A*]-(C)-[A*](U)[A*](U)[C](G)[A*](A*)[UUU](G)[A*](C)[G](U)[U](G)[U]-[A*]-[C] 3'; (B6) 5' (A)-[C']-(A)[A](C')[G](U)[C'](AAA)[U](U)[C'](G)[A](U)[A](U)[G](U) 3'; 5' [A]-(C')-[A](U)[A](U)[C'](G)[A](A)[UUU](G)[A](C')[G](U)[U](G)[U]-[A]-[C'] 3'; (B7) 5' (A*)-[C]-(A*)[A*](C)[G](U)[C](A*A*A*)[U](U)[C](G)[A*](U)[A*](U)[G](U) 3'; 5' [A*]-(C)-[A*](U)[A*](U)[C](G)[A*](A*)[UUU](G)[A*](C)[G](U)[U](G)[U]-[A*]-[C] 3'; (B8) 5' (A*)-[C]-(A*)[A*](C)[G](U)[C](A*A*A*)[U](U)[C](G)[A*](U)[A*](U)[G](U) 3'; 5' [A*]-(C)-[A*](U)[A*](U)[C](G)[A*](A*)[UUU](G)[A*](C)[G](U)[U](G)[U]-[A*]-[C] 3'; (B9) 5' (A*)-[C]-(A*)[A*](C)[G](U)[C](A*A*A*)[U](U)[C](G)[A*](U)[A*](U)[G](U) 3'; 5' [A*]-(C)-[A*](U)[A*](U)[C](G)[A*](A*)[UUU](G)[A*](C)[G](U)[U](G)[U]-[A*]-[C] 3'; (B10) 5' (A)-[C']-(A)[A](C')[G](U)[C'](AAA)[U](U)[C'](G)[A](U)[A](U)[G](U) 3'; 5' [A]-(C')-[A](U)[A](U)[C'](G)[A](A)[UUU](G)[A](C')[G](U)[U](G)[U]-[A]-[C'] 3'; (B11) 5' (A)-[C']-(A)[A](C')[G](U)[C'](AAA)[U](U)[C'](G)[A](U)[A](U)[G](U) 3'; 5' ☐ ... (B12) 5' (A)-[C']-(A)[A](C')[G](U)[C'](AAA)[U](U)[C'](G)[A](U)[A](U)[G](U) 3'; 5' ☐ ... (B13) 5' (A*)-[C']-(A*)[A*](C')[G](U)[C'](A*A *A*)[U](U)[C'](G)[A*](U)[A*](U)[G](U) 3'; 5' [A*]-(C')-[A*](U)[A*](U)[C'](G)[A*](A*)[UUU](G)[A*](C')[G](U)[UU](G)[UU]-[A*]-[C'] 3'; (B14) 5' (A*)-[C']-(A*)[A*](C')[G](U)[C'](A*A *A*)[U](U)[C'](G)[A*](U)[A*](U)[G](U) 3'; 5' [A*]-(C')-[A*](U)[A*](U)[C'](G)[A*](A*)[UUU](G)[A*](C')[G](U)[UU](G)[UU]-[A*]-[C'] 3'; (B15) 5' (A*)-[C']-(A*)[A*](C')[G](U)[C'](A*A *A*)[U](U)[C'](G)[A*](U)[A*](U)[G](U) 3'; 5' [A*]-(C')-[A*](U)[A*](U)[C'](G)[A*](A*)[UUU](G)[A*](C')[G](U)[UU](G)[UU]-[A*]-[C'] 3'; (B16) 5' (A*)-[C']-(A*)[A*](C')[G](U)[C'](A*A *A*)[U](U)[C'](G)[A*](U)[A*](U)[G](U) 3'; 5' [A*]-(C')-[A*](U)[A*](U)[C'](G)[A*](A*)[UUU](G)[A*](C')[G](U)[UU](G)[UU]-[A*]-[C'] 3'; (C1) 5' (G)-[U]-(G)[A](G)[G](C)[A](AGA)[C](C)[G](A)[A](G)[U](A)[A](A) 3'; 5' [U]-(U)-[U](A)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A)[C]-[U]-[G] 3'; (C2) 5' (G)-[U]-(G)[A](G)[G](C)[A](AGA)[C](C)[G](A)[A](G)[U](A)[A](A) 3'; 5' [U]-(U)-[U](A)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A)[C]-[U]-[G] 3'; (C3) 5' (G)-[U]-(G)[A](G)[G](C)[A](AGA)[C](C)[G](A)[A](G)[U](A)[A](A) 3'; 5' [U]-(U)-[U](A)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A)[C]-[U]-[G] 3'; (C4) 5' (G)-[U]-(G)[A](G)[G](C)[A](AGA)[C](C)[G](A)[A](G)[U](A)[A](A) 3'; 5' [U]-(U)-[U](A)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A)[C]-[U]-[G] 3'; (C5) 5' (G)-[U]-(G)[A*](G)[G](C)[A*](A*GA*)[C](C)[G](A*)[A*](G)[U](A*)[A*](A*) 3'; 5' [U]-(U)-[U](A*)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A*)[C]-[U]-[G] 3'; (C6) 5' (G)-[U]-(G)[A](G)[G](C')[A](AGA)[C'](C')[G](A)[A](G)[U](A)[A](A) 3'; 5' [U]-(U)-[U](A)[C'](U)[U](C')[G](G)[UC'U](U)[G](C')[C'](U)[C'](A)[C']-[U]-[G] 3'; (C7) 5' (G)-[U]-(G)[A*](G)[G](C)[A*](A*GA*)[C](C)[G](A*)[A*](G)[U](A*)[A*](A*) 3'; 5' [U]-(U)-[U](A*)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A*)[C]-[U]-[G] 3'; (C8) 5' (G)-[U]-(G)[A*](G)[G](C)[A*](A*GA*)[C](C)[G](A*)[A*](G)[U](A*)[A*](A*) 3'; 5' [U]-(U)-[U](A*)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A*)[C]-[U]-[G] 3'; (C9) 5' (G)-[U]-(G)[A*](G)[G](C)[A*](A*GA*)[C](C)[G](A*)[A*](G)[U](A*)[A*](A*) 3'; 5' [U]-(U)-[U](A*)[C](U)[U](C)[G](G)[UCU](U)[G](C)[C](U)[C](A*)[C]-[U]-[G] 3'; (C10) 5' (G)-[U]-(G)[A](G)[G](C')[A](AGA)[C'](C')[G](A)[A](G)[U](A)[A](A) 3'; 5' [U]-(U)-[U](A)[C'](U)[U](C')[G](G)[UC'U](U)[G](C')[C'](U)[C'](A)[C']-[U]-[G] 3'; (C11) 5' (G)-[U]-(G)[A](G)[G](C')[A](AGA)[C'](C')[G](A)[A](G)[U](A)[A](A) 3'; 5' [U]-(U)-[U](A)[C'](U)[U](C')[G](G)[UC'U](U)[G](C')[C'](U)[C'](A)[C']-[U]-[G] 3'; (C12) 5' (G)-[U]-(G)[A](G)[G](C')[A](AGA)[C'](C')[G](A)[A](G)[U](A)[A](A) 3'; 5' [U]-(U)-[U](A)[C'](U)[U](C')[G](G)[UC'U](U)[G](C')[C'](U)[C'](A)[C']-[U]-[G] 3'; (C13) 5' (G)-[U]-(G)[A*](G)[G](C')[A*](A*GA*)[C'](C')[G](A*)[A*](G)[U](A*)[A*](A*) 3'; 5' [U] - (U) - [U] (A*) [C'] (U) [U] (C') [G] (G) [UC'U] (U) [G] (C') [C'] (U) [C'] (A*) [C'] - [U] - [G] 3'; (C14) 5' (G) - [U] - (G) [A*] (G) [G] (C') [A*] (A*GA*) [C'] (C') [G] (A*) [A*] (G) [U] (A*) [A*] (A*) 3'; 5' [U] - (U) - [U] (A*) [C'] (U) [U] (C') [G] (G) [UC'U] (U) [G] (C') [C'] (U) [C'] (A*) [C'] - [U] - [G] 3'; (C15) 5' (G) - [U] - (G) [A*] (G) [G] (C') [A*] (A*GA*) [C'] (C') [G] (A*) [A*] (G) [U] (A*) [A*] (A*) 3'; 5' [U] - (U) - [U] (A*) [C'] (U) [U] (C') [G] (G) [UC'U] (U) [G] (C') [C'] (U) [C'] (A*) [C'] - [U] - [G] 3'; (C16) 5' (G) - [U] - (G) [A*] (G) [G] (C') [A*] (A*GA*) [C'] (C') [G] (A*) [A*] (G) [U] (A*) [A*] (A*) 3'; 5' [U] - (U) - [U] (A*) [C'] (U) [U] (C') [G] (G) [UC'U] (U) [G] (C') [C'] (U) [C'] (A*) [C'] - [U] - [G] 3'; (Here, A-, U-, C-, and G- represent phosphorothioate-modified ribonucleotides A, U, C, and G, respectively.) (A), (U), (C), and (G) represent ribonucleotides A, U, C, and G, respectively, that have undergone 2'-F modification. [A], [U], [C], and [G] represent ribonucleotides A, U, C, and G, respectively, that have undergone 2'-OMe modification. (A), (U), (C), and (G) represent ribonucleotides A, U, C, and G, respectively, that have undergone 2'-Ara-F modification. [A], [U], [C], and [G] represent ribonucleotides A, U, C, and G, respectively, that have undergone 2'-O-MOE modification. A*, U*, C*, and G* are each m ６ This shows ribonucleotides A, U, C, and G that have undergone A modification. A', U', C', and G' are each m ５ (This shows ribonucleotides A, U, C, and G that have undergone C modification.)

7. (1) A vector expressing a double-stranded siRNA molecule as described in claim 1, (2) A host cell having the double-stranded siRNA molecule or the vector described in claim 1, (3) A double-stranded siRNA molecule according to claim 1, or a reagent containing (1) the vector or (2) the host cell, and (4) A pharmaceutical composition containing the double-stranded siRNA molecule described in claim 1. A biological material relating to a double-stranded siRNA molecule according to claim 1, characterized in that it is one of the following.

8. The biomaterial according to claim 7, characterized in that the pharmaceutical composition further contains an androgen or an androgen receptor inhibitor.

9. The biomaterial according to claim 7, characterized in that the pharmaceutical composition contains a pharmaceutically acceptable carrier or auxiliary material.

10. The biomaterial according to claim 9, characterized in that the pharmaceutically acceptable carrier comprises microliposomes, microcells, metal particles, or polymer particles.

11. Use of a double-stranded siRNA molecule according to any one of claims 1, 3 to 5, or a composition according to claim 2 or 6, in the manufacture of a drug for preventing or treating prostate cancer.

12. Use of a biomaterial according to any one of claims 7 to 10 in the manufacture of a drug for preventing or treating prostate cancer.

13. The use according to claim 11, characterized in that the prostate cancer is advanced prostate cancer or castration-resistant prostate cancer.

14. A method for inhibiting the expression of PRDX5 and / or GPX4 genes in cells, characterized by comprising in vitro contacting cells with an effective amount of a double-stranded siRNA molecule described in any one of claims 1, 3 to 5, or a composition described in claim 2 or 6, thereby inhibiting the expression of PRDX5 and / or GPX4 genes in cells.

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