公开(公告)号：US20200071700A1

公开(公告)日：2020-03-05

申请号：US16687501

申请日：2019-11-18

Applicant: Chih-Li LIN ,  Hsin-Hua LI ,  Shi-Lung LIN ,  Te-Jen LAI

Inventor： Chih-Li LIN ,  Hsin-Hua LI ,  Shi-Lung LIN ,  Te-Jen LAI

IPC: C12N15/113 ,  C12N5/0793 ,  C12N5/10 ,  C12N15/63

Abstract: This invention generally relates to a composition and method of using recombinant microRNAs (miRNA) and their hairpin-like precursors (pre-miRNA) as therapeutic drugs for treating Alzheimer's diseases (AD). More specifically, the present invention relates to the use of man-made miRNA miR-302 precursors (pre-miR-302) for AD therapy in humans. These pre-miR-302 molecules can be mass produced in prokaryotes as a form of DNA expression-competent DNA vectors and/or hairpin-like RNAs. As prokaryotic cells do not transcribe or process hairpin-like RNAs, the present invention also teaches a method for expressing pre-miRNAs in prokaryotes, i.e. pro-miRNA, using a novel hairpin-like RNA transcription mechanism newly found in prokaryotes. Additionally, since miR-302 is a well-known embryonic stem cell (ESC)-specific factor in humans, our novel findings of this invention can be further used to advance the designs and development of novel regenerative medicine for treating many other ageing-related degenerative diseases, such as Parkinson's diseases, osteoporosis, diabetes, and cancers.

公开(公告)号：US20190085335A1

公开(公告)日：2019-03-21

申请号：US16135723

申请日：2018-09-19

Applicant: Shi-Lung LIN ,  Samantha CHANG-LIN ,  Donald CHANG

Inventor： Shi-Lung LIN ,  Samantha CHANG-LIN ,  Donald CHANG

IPC: C12N15/113 ,  C12N15/70 ,  C12N15/11 ,  C12P19/34

Abstract: This invention generally relates to a composition and its method of use for inducing adult stem cell (ASC) expansion and/or derivation in vitro, using miR-302-like pre-miRNAs, shRNAs and/or siRNAs, all of which contain a shared sequence of 5′-UAAGUGCUUC CAUGUUU-3′ (SEQ ID NO: 7) in the 5′-end, and further in conjunction with the use of some wound-healing-related defined factors, including but not limited to basic fibroblast growth factor (bFGF)/fibroblast growth factor 2 (FGF-2), leukemia inhibitory factor (LIF), insulin-like growth factor (IGF), Epidermal growth factor (EGF), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), transforming growth factor (TGF), tumor necrosis factor (TNF), stem cell factor (SCF), homeobox proteins (HOX), Notch, GSK, Wnt/beta-Catenin signals, interleukins, and/or bone morphogenetic proteins (BMPs). The principle of the present invention is related to a novel mechanism of inducible symmetric ASC division recently found in a skin wound healing model in vivo. The resulting amplified ASCs are useful for treating a variety of human aging- and cell dysfunction-associated disorders, including but not limited to Alzheimer's disease, Parkinson's disease, motor neuron disease, stroke, diabetes, osteoporosis, myocardial infraction, hemophilia, anemia, AIDS, leukemia, lymphoma and many kinds of cancers as well as aging.

公开(公告)号：US20250019709A1

公开(公告)日：2025-01-16

申请号：US18795444

申请日：2024-08-06

Applicant: Shi-Lung LIN ,  Sam LIN ,  Wei-Chih HUNG ,  Hsien-Lin LIU ,  Yu-Jing WU

Inventor： Shi-Lung LIN ,  Sam LIN ,  Wei-Chih HUNG ,  Hsien-Lin LIU ,  Yu-Jing WU

IPC: C12N15/113

Abstract: This invention relates to a novel modified RNA composition (called oxo-RNA) comprising at least an oxo-nucleotide (containing oxopurine) in its 3′-end region (e.g. 3′-tail, 3′-UTR). The oxo-nucleotide includes 8-hydroxyguanine/8-oxoguanine/7,8-dihydro-8-oxoguanine (or called oxo-G/oxo-dG) and 8-hydroxyadenine/8-oxoadenine (or called oxo-A/oxo-dA). Oxo-RNA can be a single-stranded RNA sequence or double-stranded duplex, or even an RNA-DNA hybrid duplex. Advantageously, this new oxo-RNA composition not only greatly enhance RNA/DNA stability and functionality but also can prevent TREX1-mediated degradation and the related non-specific immunity over-activation (e.g. cytokine storm). Most importantly, the constructs of oxo-RNA can be designed to mimic antisense RNA oligonucleotide (aRNA-ASO), small interfering RNA (siRNA), short hairpin RNA (shRNA), microRNA (miRNA) mimic, microRNA precursor (pre-miRNA), double-stranded RNA (dsRNA), RNA-DNA hybrid, long noncoding RNA (IncRNA), small activating RNA (saRNA), messenger RNA (mRNA), and/or self-amplifying RNA/mRNA (saRNA/samRNA), or a combination thereof.

公开(公告)号：US20240093286A1

公开(公告)日：2024-03-21

申请号：US18479983

申请日：2023-10-03

Applicant: Shi-Lung LIN ,  Sam Lin

Inventor： Shi-Lung LIN ,  Sam Lin

IPC: C12Q1/686 ,  A61K31/713 ,  C12N15/10

CPC classification number: C12Q1/686 ,  A61K31/713 ,  C12N15/1096

Abstract: This invention relates to a novel composition of RNA/mRNA medicines and/or vaccines produced by using Replicase/RNA-dependent RNA polymerase (RdRP)-mediated RNA Cycling Reaction (RCR). This RCR-amplifiable RNA/mRNA composition comprises at least a replicase/RdRP-binding site (RdRP-BS) in the 5′-end or 3′-end, or both, of a desired RNA sequence of interest, to form a self-amplifying RNA/mRNA (samRNA) platform. The samRNA platform so obtained is useful for designing and developing a variety of self-amplifying RNA/mRNA (samRNA) constructs, of which the desired RNA sequences may include, but not limited to, antisense oligonucleotide RNA (aRNA; ASO), small interfering RNA (siRNA), short hairpin RNA (shRNA), microRNA (miRNA)/miRNA precursor (pre-miRNA), long non-coding RNA (lncRNA), and/or messenger RNA (mRNA), or a combination thereof. The present RdRP-BS designs in said samRNA are derived or modified from the identified RdRP-BS motifs of coronavirus (e.g. SARS-CoV-2-associated viruses) and/or hepatitis C virus (HCV) in either single-stranded or double-stranded conformation, or a combination thereof.

公开(公告)号：US20200318110A1

公开(公告)日：2020-10-08

申请号：US16305030

申请日：2017-02-24

Applicant: Shi-Lung LIN ,  David TS WU

Inventor： Shi-Lung LIN ,  David TS WU ,  Hsuan-Hsuan LU

IPC: C12N15/113 ,  A61K31/7105 ,  A61P35/04

Abstract: This invention generally relates to a composition and method of using man-made small RNAs, such as small interfering RNAs (siRNA), microRNAs (miRNA) and their hairpin-like precursors (pre-miRNA), as tumor suppressing anti-cancer drugs for treating human tumors and cancers, in particular, but not limited, for treating skin (melanoma), blood (leukemia), prostate, breast, liver and lung cancers as well as various neoplastic tumors, such as brain tumors and teratocarcinomas that contain a variety of tumorous and cancerous cells derived from all three germ layers of tissues, including ectoderm, mesoderm and endoderm. More specifically, the present invention relates to the use of miR-302-like siRNA (siR-302) and/or miR-302 precursors (pre-miR-302) for developing novel medicines and therapies against a variety of human cancers, in particular lung cancers.

公开(公告)号：US20160220525A1

公开(公告)日：2016-08-04

申请号：US15074321

申请日：2016-03-18

Applicant: Shi-Lung LIN ,  Samantha CHANG-LIN ,  Yi-Wen LIN ,  Donald CHANG

Inventor： Shi-Lung LIN ,  Samantha CHANG-LIN ,  Yi-Wen LIN ,  Donald CHANG

IPC: A61K31/221 ,  A61K9/08 ,  A61K31/7024 ,  A61K9/00

CPC classification number: A61K31/221 ,  A61K9/0053 ,  A61K9/08 ,  A61K31/7012 ,  A61K31/7024 ,  C12N15/111 ,  C12N15/113 ,  C12N2310/141 ,  C12N2320/32 ,  C12N2330/50 ,  C12N2330/51

Abstract: This invention is related to a novel sugar-like chemical composition and its use for diabetes therapy. Particularly, the present invention teaches the use of monosaccharide-like glycylated sugar alcohol compounds to block or reduce sugar absorption in diabetes patients, so as to prevent the risk of hyperglycemia symptoms. Glycylation of sugar alcohols is a totally novel reaction that has never been reported before. Therefore, the novelty of the present invention is that for the first time glycylated sugar alcohols not only was found but also was found to be useful for treating Diabetes mellitus. In addition, the present invention teaches a method for producing these glycylated sugar alcohols. In sum, the present invention includes not only a kind of novel sugar-like chemical compositions and its use for treating diabetes but also a state-of-the-art protocol and methodology for producing such a novel composition via glycylation of sugars and sugar alcohols.

Abstract translation: 本发明涉及新型糖类化学组合物及其用于糖尿病治疗的用途。 特别地，本发明教导了使用单糖样的糖化糖醇化合物来阻止或减少糖尿病患者的糖吸收，以防止高血糖症状的风险。 糖醇的糖基化是一种完全新颖的反应，从未在之前报道过。 因此，本发明的新颖性不仅首次发现了糖化糖醇，而且被发现可用于治疗糖尿病。 此外，本发明教导了一种制备这些糖基化糖醇的方法。 总之，本发明不仅包括一种新型的糖类化学组合物及其用于治疗糖尿病的用途，而且还包括通过糖和糖醇的糖基化制备这种新型组合物的最新方案和方法 。

公开(公告)号：US20230099592A1

公开(公告)日：2023-03-30

申请号：US17930292

申请日：2022-09-07

Applicant: Shi-Lung LIN ,  Sam Lin ,  Chun-Hung Lin

Inventor： Shi-Lung LIN ,  Sam Lin ,  Chun-Hung Lin

IPC: C12N15/10 ,  A61P31/12

Abstract: This invention relates to a novel composition and method for RNA/mRNA production as well as amplification using viral RNA replicase and/or RNA-dependent RNA polymerase (RdRp) enzymes and the use of associated RNA/mRNA products thereof. The present invention can be used for manufacturing and amplifying all varieties of RNA/mRNA sequences carrying at least a replicase/RdRp-binding site in the 5′- or 3′-end, or both. The RNA/mRNA so obtained is useful for not only producing mRNA vaccines and/or RNA-based medicines but for generating the mRNA-associated proteins, peptides, and/or antibodies under an in-vitro as well as in-cell translation condition. Principally, the present invention is a novel RNA replicase/RdRp-mediated RNA/mRNA amplification method, namely Replicase Cycling Reaction (RCR). The RNA replicases involved in RCR include but not limited to viral and/or bacteriophage RNA-dependent RNA polymerases (RdRp) in either modified or non-modified mRNA and/or protein compositions, particularly coronaviral (e.g. COVID-19) and hepatitis C viral (HCV) RdRp enzymes.

公开(公告)号：US20160264974A1

公开(公告)日：2016-09-15

申请号：US15167226

申请日：2016-05-27

Applicant: Shi-Lung LIN ,  Donald CHANG ,  David TS WU

Inventor： Shi-Lung LIN ,  Donald CHANG ,  David TS WU

IPC: C12N15/113 ,  C12P19/34 ,  C12N15/70

CPC classification number: C12N15/1135 ,  C12N15/111 ,  C12N15/113 ,  C12N15/70 ,  C12N2310/14 ,  C12N2310/141 ,  C12N2320/30 ,  C12N2330/50 ,  C12N2330/51 ,  C12P19/34

Abstract: This invention generally relates to a composition and its production method useful for developing drugs/vaccines and/or therapies against a variety of degenerative diseases in humans. Particularly, the present invention teaches the essential steps of production and purification processes necessary for producing small hairpin-like RNA (shRNA) compositions, such as microRNA precursors (pre-miRNA) and short interfering RNAs (siRNA), which are useful for treating human ageing related diseases, such as, but not limited, Alzheimer's diseases, Parkinson's diseases, osteoporosis, diabetes, and cancers. The novelty of the present invention is to create an artificially enhanced adaptation environment for prokaryotic cells to adopt eukaryotic pol-2 and/or pol-2-like promoters for transcribing desired ncRNAs and/or their precursors without going through error-prone prokaryotic promoters, so as to improve the productive efficiency and reading fidelity of the shRNA transcription in the prokaryotic cells. The resulting shRNAs, preferably pre-miRNAs and siRNAs, are useful for developing therapeutic drugs against human degenerative diseases, particularly through a mechanism to induce CD34-positive stem cell expansion and/or regeneration.

Abstract translation: 本发明一般涉及用于开发针对人类各种退行性疾病的药物/疫苗和/或疗法的组合物及其制备方法。 特别地，本发明教导了用于生产小发夹样RNA（shRNA）组合物，例如可用于治疗人类的微小RNA前体（pre-miRNA）和短干扰RNA（siRNA））所需的生产和纯化过程的基本步骤 衰老相关疾病，例如但不限于阿尔茨海默病，帕金森病，骨质疏松症，糖尿病和癌症。 本发明的新颖之处在于为原核细胞产生人造增强的适应环境，以采用真核的pol-2和/或pol-2样启动子来转录所需的ncRNA和/或其前体而不经历易错的原核启动子， 从而提高原核细胞中shRNA转录的生产效率和阅读保真度。 产生的shRNA，优选pre-miRNAs和siRNAs可用于开发针对人退行性疾病的治疗药物，特别是通过诱导CD34阳性干细胞扩增和/或再生的机制。

公开(公告)号：US20220411848A1

公开(公告)日：2022-12-29

申请号：US17648336

申请日：2022-01-19

Applicant: Shi-Lung LIN ,  Sam LIN ,  Chun-Hung LIN

Inventor： Shi-Lung LIN ,  Sam LIN ,  Chun-Hung LIN

IPC: C12Q1/686 ,  C12N9/12

Abstract: This invention generally relates to a novel RNA/mRNA production and amplification method using viral RNA replicase and/or RNA-dependent RNA polymerase (RdRp) enzymes as well as the associated mRNAs thereof. The present invention can be used for manufacturing and amplifying all varieties of RNA/mRNA sequences carrying at least an RdRp-binding site in the 5′- or 3′-end, or both. The RNA/mRNA so obtained is useful for not only producing mRNA vaccines and/or RNA-based medicines but also for generating the mRNA-associated proteins, peptides, and/or antibodies under an in-vitro as well as in-cell translation condition. Principally, the present invention is a novel RNA replicase-mediated RNA/mRNA amplification method, namely Replicase Cycling Reaction (RCR). The RNA replicases involved in RCR include but not limited to viral and/or bacteriophage RNA-dependent RNA polymerases (RdRp), particularly coronaviral and hepatitis C viral (HCV) RdRp enzymes.

公开(公告)号：US20220396798A1

公开(公告)日：2022-12-15

申请号：US17489357

申请日：2021-09-29

Applicant: Shi-Lung LIN ,  Samantha CHANG-LIN ,  Jack SK CHEN ,  David TS WU ,  Chia-Ning SHEN ,  Mei-Jung WANG

Inventor： Shi-Lung LIN ,  Samantha CHANG-LIN ,  Jack SK CHEN ,  David TS WU ,  Chia-Ning SHEN ,  Mei-Jung WANG

IPC: C12N15/113 ,  C12N15/85

Abstract: This invention relates to a novel mRNA composition and its production method useful for developing and manufacturing RNA-based anti-viral and/or anti-cancer vaccines and medicines. This invention includes two types of mRNA constructs, namely “5′-hairpin messenger RNA (5hmRNA)” and “messenger-hairpin-messenger RNA (mhmRNA)”, respectively. Both of 5hmRNA and mhmRNA contain at least a hairpin-like stem-loop RNA structure. The 5hmRNA contains at least a stem-loop RNA structure in the 5′-UTR of a protein/peptide-coding mRNA, while the mhmRNA contains a middle stem-loop structure flanked with two protein/peptide-coding mRNA sequences on both sides. In mhmRNA, the first 5′-mRNA preferably encodes an RNA replicase, for amplifying the second 3′-mRNA in transfected cells. After transfection into target cells, 5hmRNA and mhmRNA can be further translated into at least a desired protein/peptide. To produce highly structured 5hmRNA and mhmRNA, a novel PCR-IVT methodology has been developed and used with a specially designed RNA polymerase-helicase mixture reaction