PHARMACEUTICAL COMPOSITION COMPRISING THE AAT RNAi DRUG SUBSTANCE AND USES OF THE AAT RNAi DRUG SUBSTANCE AND THE PHARMACEUTICAL COMPOSITION COMPRISING SAID SUBSTANCE FOR TREATING ALPHA-1 ANTITRYPSIN DEFICIENCY (AATD)

Description

1 / 46 “PHARMACEUTICAL COMPOSITION COMPRISING THE AAT RNAi DRUG SUBSTANCE AND USES OF THE AAT RNAi DRUG SUBSTANCE AND THE PHARMACEUTICAL COMPOSITION COMPRISING SAID SUBSTANCE FOR TREATING ALPHA-1 ANTITRYPSIN DEFICIENCY (AATD)” Divided from BR 11 2021 024534 0, deposited on 05 / 06 / 2020 Sequence Listing

[001] This request contains a Sequence Listing, which was submitted in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy is named 30674_WO1_SequenceListing.txt and is 6 kb in size. Field of invention

[002] Methods for treating alpha-1 antitrypsin deficiency (AATD) in a human subject are disclosed here, including the treatment of symptoms and diseases caused by AATD, using pharmaceutical compositions that include RNA interference agents (RNAi) that inhibit alpha-1 antitrypsin gene expression. Foundation

[003] Alpha-1 antitrypsin (AAT, α1-antitrypsin or A1AT) is a protease inhibitor belonging to the serpin superfamily encoded in humans by the SERPINA1 gene. Normal AAT protein is a circulating glycoprotein protease inhibitor synthesized primarily in the liver by hepatocytes and secreted into the blood. The known physiological function of AAT is to inhibit neutrophil proteases, which serve to protect host tissues from nonspecific injury during periods of inflammation.

[004] Alpha-1 antitrypsin deficiency (AATD) is an autosomal codominant genetic disorder that results in low circulating levels of AAT and causes early lung disease in adults and liver disease in children and adults. A Petition 870260074731, dated 07 / 27 / 2026, page 13 / 97 The prevalence range of AAT deficiency (AATD) is approximately 1 in 1,500 to 5,000 individuals and generally affects people of European descent.

[005] The most clinically significant form of AATD is caused by the Z mutation. The mutant Z allele, through a single point mutation, makes the mutant Z-form AAT protein (the “Z-AAT protein”) prone to abnormal folding, causing intracellular retention in the endoplasmic reticulum (ER) of hepatocytes. Other rarer mutations also result in accumulated misfolded protein in hepatocytes. The mutant Z-AAT protein monomers are able to accumulate into polymer aggregates, which are sometimes called “globules.” The masses of polymeric globules stress the ER and trigger a cycle of continuous hepatocyte injury and healing, leading to fibrosis, cirrhosis, and an increased risk of hepatocellular carcinoma.Furthermore, the absence of circulating antiprotease activity leaves the lung vulnerable to neutrophil elastase damage, particularly in the context of pulmonary inflammation, resulting in the development of respiratory complications such as emphysema or other lung disease.

[006] Individuals with the homozygous PiZZ genotype exhibit severe functional AAT deficiency. Weekly use of AAT augmentation therapy, using purified human AAT, helps prevent lung damage in affected individuals. Currently marketed products include, for example, Prolastin®-C, Prolastin®, Glassia™, Aralast® NP, and Zemaira®. However, while administration of purified AAT may improve or help prevent lung damage caused by the absence or low levels of endogenously secreted AAT, AATD patients (with an AAT mutation resulting in polymer formation) remain vulnerable to endoplasmic reticulum liver storage disease caused by the deposition and accumulation of excessive abnormally folded AAT protein. The Z-AAT protein accumulated in a “globule” conformation in hepatocytes is a well-known histological feature of AATD liver disease. Petition 870260074731, dated 07 / 27 / 2026, page 14 / 97 3 / 46 and is believed to lead to proteotoxic effects responsible for inducing liver injury, including liver cell injury and death and chronic liver injury in individuals with AATD (see, for example, D. Lindblad et al., Hepatology 2007, 46: 1228-1235). It has been reported that null / null patients, who do not produce AAT, develop severe lung disease but have normal liver morphology, providing evidence that the accumulation of mutant AAT, and not the lack of circulating AAT, leads to liver disease (Feldman, G. et al, The Ultrastructure of Hepatocytes in alpha-1 antitrypsin deficiency with genotype Pi_, Gut. 1975; 16:796-799).

[007] AATD predisposes individuals to liver disease in children and adults and to early-onset emphysema in adults. Patients with AATD frequently develop liver disease, which can be severe or fatal, even in childhood. While some patients with AATD initially escape detection, eventually fibrosis accumulates and leads to clinically apparent liver disease. Clinical presentations of liver injury include chronic hepatitis, cirrhosis, increased risk of hepatocellular carcinoma, transaminitis, cholestasis, fibrosis, and even fulminant hepatic failure.

[008] The accumulation of Z-AAT protein globules in hepatocytes has been clearly identified as the cause of progressive liver disease in patients with AATD. Elimination of the mutant protein accumulation in hepatocytes may halt the progression of liver disease. Removal of the mutant protein insult may also allow regression of already present fibrosis. Currently, there is no clinically approved treatment to prevent the onset, delay the progression, or treat liver disease caused by AATD.

[009] RNAi agents have emerged as a promising avenue for the treatment of patients with AATD. Dosing strategies are an important consideration in the treatment of AATD with RNAi agents. Less frequent doses are valued by patients, leading to greater adherence and lower doses. Petition 870260074731, dated 07 / 27 / 2026, page 15 / 97 4 / 46 smaller doses may be advantageous in the overall safety profile of the drug. Therefore, there is a need for a low-dose, infrequent method for the treatment of AATD. Summary

[010] This document describes methods for treating alpha-1 antitrypsin deficiency (AATD) in a human subject in need thereof. In one aspect, the methods comprise administering to the human subject a pharmaceutical composition that includes the composition described in Table 2 (i.e., AAT RNAi Drug Substance, also referred to in this document as ADS001) at a dose between about 5 mg and about 300 mg of the AAT RNAi Drug Substance, wherein the pharmaceutical composition is administered subcutaneously and there is at least about one month between doses (i.e., at least monthly dosing). In some embodiments, the pharmaceutical composition used in the methods disclosed in this document comprises, consists of, or essentially consists of the formulated AAT RNAi Drug Substance as described in Table 3 (also referred to in this document as ADS-001-1).

[011] In addition, methods of treating AATD in a human subject in need thereof are described in this document, the methods comprising administering to the human subject a pharmaceutical composition that includes the AAT RNAi Drug Substance as described in Table 2 (i.e., ADS001) at a dose of between about 5 mg and about 200 mg, wherein the pharmaceutical composition is administered subcutaneously and there is at least about one month between dose administrations (i.e., monthly dosing).

[012] In addition, methods of treating AATD in a human subject in need thereof are described in this document, the methods comprising administering to the human subject a pharmaceutical composition that includes the AAT RNAi Drug Substance as described in Table 2 (i.e., ADS). Petition 870260074731, dated 07 / 27 / 2026, page 16 / 97 5 / 46 001) at a dose of between approximately 5 mg and approximately 300 mg, where the pharmaceutical composition is administered subcutaneously and there are approximately three months between dose administrations (i.e., quarterly dosing).

[013] Also described in this document are methods of treating AATD in a human subject in need thereof, the methods comprising administering to the human subject a pharmaceutical composition which includes the AAT RNAi Drug Substance as described in Table 2 (i.e., ADS001) at a dose of between about 5 mg and about 200 mg, wherein the pharmaceutical composition is administered subcutaneously and there are about three months between dose administrations (i.e., quarterly dosing).

[014] This document describes methods of treating AATD in a human subject in need thereof, the methods comprising administering to the human subject a pharmaceutical composition that includes the AAT RNAi Drug Substance as described in Table 2 (i.e., ADS-001) at a dose of between about 5 mg and about 300 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein the initial dose is followed by a second dose about one month later and, subsequently, for subsequent doses, there are about three months between dose administrations.

[015] This document describes methods of treating AATD in a human subject in need thereof, the methods comprising administering to the human subject a pharmaceutical composition that includes the AAT RNAi Drug Substance as described in Table 2 (i.e., ADS-001) at a dose of between about 5 mg and about 200 mg, wherein the pharmaceutical composition is administered subcutaneously, and wherein the initial dose is followed by a second dose about one month later and, subsequently, for subsequent doses, there are about three months between dose administrations.

[016] In some modalities, the dose of AAT RNAi Drug Substance Petition 870260074731, dated 07 / 27 / 2026, p. 17 / 97 6 / 46 administered in each dose is between approximately 25 mg and approximately 200 mg. In some embodiments, the dose of AAT RNAi Drug Substance administered in each dose is between approximately 100 mg and approximately 200 mg. In some embodiments, the dose of AAT RNAi Drug Substance administered in each dose is approximately 100 mg. In some embodiments, the dose of AAT RNAi Drug Substance administered in each dose is approximately 200 mg. In some embodiments, the dose of AAT RNAi Drug Substance administered in each dose does not exceed 200 mg.

[017] The treatment methods disclosed herein may slow or halt the progression of liver disease in a human subject with AATD, which may allow for the repair of fibrotic tissue. The methods disclosed herein may, in some modalities, treat AATD liver diseases, including fibrosis, cirrhosis, increased risk of hepatocellular carcinoma, chronic hepatitis, transaminitis, cholestasis, fulminant hepatic failure, and other liver-related conditions and diseases caused by AATD.

[018] The pharmaceutical compositions that include AAT RNAi agents disclosed herein may be administered to a human subject to inhibit alpha-1 antitrypsin gene expression in the subject. In some embodiments, the subject is a human who has been previously diagnosed with AATD.

[019] Another aspect of the invention provides for utilizing the AAT RNAi Drug Substance described in Table 2 for the treatment of alpha-1 antitrypsin deficiency (AATD) in a human subject in need thereof, wherein the use comprises administering to the patient a pharmaceutical composition comprising the AAT RNAi Drug Substance described in Table 2 at a dose of between about 5 mg and about 300 mg of the AAT RNAi Drug Substance, wherein the pharmaceutical composition is administered once a month by subcutaneous injection.

[020] Another aspect of the invention provides the use of the RNAi Drug Substance Petition 870260074731, dated 07 / 27 / 2026, p. 18 / 97 7 / 46 of AAT described in Table 2 for the treatment of alpha-1 antitrypsin deficiency (AATD) in a human subject in need thereof, wherein the use comprises administering to the patient a pharmaceutical composition comprising the AAT RNAi Drug Substance described in Table 2 at a dose of between about 5 mg and about 300 mg of the AAT RNAi Drug Substance, wherein the pharmaceutical composition is administered once every three months by subcutaneous injection.

[021] Other objects, features, aspects and advantages of the invention will be evident from the following detailed description, attached figures and claims. Brief Description of the Drawings

[022] FIG. 1A to 1E. Representation of the chemical structure of the AAT RNAi Drug Substance described in Table 2 (herein referred to as ADS-001; i.e., AAT RNAi agent conjugated to a tridentate N-acetylgalactosamine targeting group at the 5' terminal end of the sense strand), shown in the form of a sodium salt.

[023] FIG. 2A to 2E. Representation of the chemical structure of the AAT RNAi Drug Substance described in Table 2, shown in a free acid form.

[024] FIG. 3. Schematic diagram of the modified sense and antisense strands of the AAT RNAi Drug Substance described in Table 2 (hereinafter referred to as ADS-001; i.e., AAT RNAi agent conjugated to a tridentate N-acetylgalactosamine targeting group at the 5' terminal end of the sense strand). The following abbreviations are used in Figure 3: a, c, gu are nucleotides modified with 2'-O-methyl; Af, Cf, Gf and Uf are nucleotides modified with 2'-fluoro (also referred to in the art as 2'-deoxy-2'-fluoro); o is a phosphodiester linkage; s is a phosphorothioate linkage; invAb is an inverted abasic residue or subunit; (NAG37)s is a tridentate N-acetylgalactosamine targeting ligand having the following chemical structure: Petition 870260074731, dated 07 / 27 / 2026, p. 19 / 97 8 / 46 (shown in the form of sodium salt), OH (shown in the form of free acid).

[025] FIG. 4. Final Phase I study design and dose escalation schedule for the Phase I clinical trial described in Example 2.

[026] FIG. 5. Graph showing serum AAT levels in healthy human volunteers (NHV) administered placebo (all cohorts) or 35 mg of AAT RNAi Drug Substance (Cohort 1) from the Phase I clinical trial described in Example 2. As shown in Figures 5 to 11, “Active” refers to the nucleotide-modified AAT RNAi described in Table 2 (administered as Petition 870260074731, dated 07 / 27 / 2026, page 20 / 97 9 / 46 modified nucleotides AAT RNAi formulated as described in Table 3).

[027] FIG. 6. Graph showing serum AAT levels in NHVs administered with placebo (all cohorts) or a single 100 mg dose of modified AAT RNAi nucleotides (Cohort 2b) from the Phase I clinical trial described in Example 2.

[028] FIG. 7. Graph showing serum AAT levels in NHVs administered with placebo (all cohorts) or a single 200 mg dose of modified AAT RNAi nucleotides (Cohort 3b) from the Phase I clinical trial described in Example 2.

[029] FIG. 8. Graph showing serum AAT levels in NHVs administered with placebo (all cohorts) or a single 300 mg dose of modified AAT RNAi nucleotides (Cohort 4b) from the Phase I clinical trial described in Example 2.

[030] FIG. 9. Graph showing serum AAT levels in NHVs administered with placebo (all cohorts) or three 100 mg doses of modified AAT RNAi nucleotides administered monthly (Cohort 2) from the Phase I clinical trial described in Example 2.

[031] FIG. 10. Graph showing serum AAT levels in NHVs administered with placebo (all cohorts) or three doses of 200 mg of modified AAT RNAi nucleotides administered monthly (Cohort 3) from the Phase I clinical trial described in Example 2.

[032] FIG. 11. Graph showing serum AAT levels in NHVs administered with placebo (all cohorts) or three doses of 300 mg of modified AAT RNAi nucleotides administered monthly (Cohort 4) from the Phase I clinical trial described in Example 2. Detailed Description RNAi agents Petition 870260074731, dated 07 / 27 / 2026, page 21 / 97 10 / 46

[033] The methods described in this document include administering a pharmaceutical composition to a human subject, wherein the pharmaceutical composition includes a composition containing an RNA interference agent (RNAi) (referred to in this document and in the art as an RNAi agent or an RNAi trigger) capable of inhibiting the expression of an AAT gene. In some embodiments, the methods described in this document include administering a pharmaceutical composition to a human subject, wherein the pharmaceutical composition includes the AAT RNAi Drug Substance described in Table 2 (also referred to as ADS001). Compositions suitable for use in the methods disclosed herein consist of an RNAi agent that inhibits the expression of an AAT gene in a human subject and a targeting fraction or group.In some embodiments, the RNAi agent includes the nucleotide sequences provided in Tables 1A and 1B, and the sense strand of the RNAi agent is further linked or conjugated to a targeting group comprising three N-acetylgalactosamine targeting moieties (see, for example, Table B). An RNAi agent that inhibits the expression of an AAT gene in a human subject is referred to as an “AAT RNAi agent”.

[034] In general, AAT RNAi agents comprise a sense strand (also referred to as a passenger strand) and an antisense strand (also referred to as a guide strand) that are annealed to form a duplex. The AAT RNAi agents disclosed herein include an RNA or RNA-like oligonucleotide molecule (e.g., chemically modified RNA) capable of degrading or inhibiting the translation of AAT mRNA messenger RNA (mRNA) transcripts in a sequence-specific manner. The AAT RNAi agents disclosed in this document may operate via the RNA interference mechanism (i.e., inducing RNA interference through interaction with the RNA interference pathway machinery (RNA-induced silencing complex or RISC) of cells). Petition 870260074731, dated 07 / 27 / 2026, p. 22 / 97 11 / 46 mammals), or by any alternative mechanism(s) or pathway(s). Although AAT RNAi agents, as that term is used in this document, are believed to operate primarily through the RNA interference mechanism, the disclosed RNAi agents are not linked to or limited to any particular pathway or mechanism of action. RNAi agents generally consist of a sense strand and an antisense strand, each 16 to 49 nucleotides long, and include, but are not limited to: short or small interfering RNAs (siRNAs), double-stranded RNAs (dsRNAs), microRNAs (miRNAs), short hook RNAs (shRNAs), and dicer substrates.

[035] The length of a sense strand of AAT RNAi agent is typically 16 to 49 nucleotides long, and the length of an antisense strand of AAT RNAi agent is typically 18 to 49 nucleotides long. In some embodiments, the sense and antisense strands are independently 17 to 26 nucleotides long. In some embodiments, the sense and antisense strands are independently 21 to 26 nucleotides long. In some embodiments, the sense and antisense strands are independently 21 to 24 nucleotides long. In some embodiments, the sense and antisense strands independently have lengths of 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the sense and antisense strands both have lengths of 21 nucleotides. The sense and antisense strands may have the same length or different lengths.The sense and antisense strands can also form pendant nucleotides at one or both ends of the AAT RNAi agent.

[036] AAT RNAi agents inhibit, silence, or knockdown the expression of the AAT gene. As used in this document, the terms “silence,” “reduce,” “inhibit,” “downregulate,” or “knockdown,” when referring to AAT expression, mean that gene expression, as measured by RNA level, is affected. Petition 870260074731, dated 07 / 27 / 2026, p. 23 / 97 12 / 46 The transcribed gene or the level of polypeptide, protein, or protein subunit translated from mRNA in a cell, cell group, tissue, organ, or subject in which the gene is transcribed is reduced when the cell, cell group, tissue, organ, or subject is treated with the RNAi agent compared to a second cell, cell group, tissue, organ, or subject that has not been treated. In some cases, the reduction in gene expression is measured by comparing baseline levels of AAT mRNA or AAT protein in a human subject before administration of a composition comprising an AAT RNAi agent with levels of AAT mRNA or AAT protein after administration of the therapeutic.

[037] Inhibition, silencing, or knockdown of the AAT gene can be measured by any appropriate assay or method known in the art. The non-limiting examples presented in this document, as well as the examples presented in International Patent Application Publication WO 2018 / 132432 (Patent Application PCT / US2018 / 013102), which is incorporated by reference herein in its entirety, provides certain examples of appropriate assays for measuring the inhibition of AAT gene expression. A reference AAT mRNA gene transcript (SERPINA1) for normal humans (referred to as transcript variant 1; GenBank NM_000295.4) can be found at SEQ ID NO: 1.

[038] Suitable AAT RNAi agents for use in the methods disclosed herein may be covalently linked or conjugated to a targeting group that includes one or more N-acetylgalactosamine moieties. In embodiments, suitable AAT RNAi agents for use in the methods disclosed in this document are covalently linked or conjugated to a targeting group that includes one or more N-acetylgalactosamine moieties, thus forming the AAT RNAi Drug Substance described in Table 2. In some embodiments, the methods described in this document include the administration of the AAT RNAi Drug Substance described in Table 2. The RNAi Drug Substance Petition 870260074731, dated 07 / 27 / 2026, p. 24 / 97 13 / 46 of AAT described in Table 2 includes the AAT RNAi agent shown in Table 1A (antisense strand) and Table 1B (sense strand). The N-acetylgalactosamine moieties facilitate the targeting of the AAT RNAi agent to readily present asialoglycoprotein receptors (ASGPr) on the surface of hepatocytes, leading to the internalization of the AAT RNAi agent by endocytosis or other means.

[039] AAT RNAi agents that may be suitable for use in the methods disclosed in this document include an antisense strand that has a complementarity region with at least one portion of an AAT mRNA. AAT RNAi agents and AAT RNAi Drug Substances suitable for use in the disclosed methods are described in International Patent Application Publication WO 2018 / 132432 (PCT / US2018 / 013102 Patent Application), which as noted above is incorporated by reference herein in its entirety.

[040] As used in this document, the terms “sequence” and “nucleotide sequence” mean a succession or order of nucleobases or nucleotides, described with a succession of letters using standard nomenclature. As used in this document, the terms “nucleobase” and “nucleotide” have the same meaning as commonly understood in the art.

[041] As used in this document, the term “complementary,” when used to describe a first nucleotide sequence (e.g., antisense strand of RNAi agent) in relation to a second nucleotide sequence (e.g., sense strand of RNAi agent or targeted mRNA sequence), means the ability of an oligonucleotide comprising the first nucleotide sequence to hybridize (form base-pair hydrogen bonds under mammalian physiological conditions (or otherwise suitable conditions) and form a duplex or double helix structure under certain standard conditions) with an oligonucleotide comprising the second nucleotide sequence. One skilled in the art would be able to select the most appropriate set of conditions for a Petition 870260074731, dated 07 / 27 / 2026, page 25 / 97 14 / 46 Hybridization test. Complementary sequences include Watson-Crick base pairs or non-Watson-Crick base pairs and include natural or modified nucleotides or nucleotide mimics, at least to the extent that the above hybridization requirements are met. Sequence identity or complementarity is independent of modification. For example, ae Af, as defined in this document, are complementary to U (or T) and identical to A for purposes of determining identity or complementarity.

[042] As used in this document, “perfectly complementary” or “fully complementary” means that all (100%) of the bases in a contiguous sequence of a first oligonucleotide will hybridize with the same number of nucleotides in a contiguous sequence of a second oligonucleotide. The contiguous sequence may comprise all or part of a first or second nucleotide sequence.

[043] As used in this document, “partially complementary” means that in a hybridized pair of nucleotide sequences, at least 70%, but not all, of the bases in a contiguous sequence of a first oligonucleotide will hybridize with the same number of bases in a contiguous sequence of a second polynucleotide.

[044] As used in this document, “substantially complementary” means that in a hybridized pair of nucleotide sequences, at least 85%, but not all, of the bases in a contiguous sequence of a first oligonucleotide will hybridize with the same number of bases in a contiguous sequence of a second polynucleotide. The terms “complementary”, “fully complementary”, “partially complementary”, and “substantially complementary” in this document are used with respect to the nucleotide matching between the sense strand and the antisense strand of an RNAi agent, or between the antisense strand of an RNAi agent and a sequence of an AAT mRNA. Petition 870260074731, dated 07 / 27 / 2026, p. 26 / 97 15 / 46

[045] As used in this document, the term “substantially identical” or “substantially identical” applied to nucleic acid sequences means that a nucleic acid sequence comprises a sequence that has at least about 85% sequence identity or more, for example, at least 90%, at least 95%, or at least 99% identity, compared to a reference sequence. The percentage of sequence identity is determined by comparing two optimally aligned sequences in a comparison window. The percentage is calculated by determining the number of positions where the identical nucleic acid base occurs in both sequences to reach the number of paired positions, dividing the number of paired positions by the total number of positions in the comparison window, and multiplying the result by 100 to reach the percentage of sequence identity.The inventions disclosed herein involve nucleotide sequences substantially identical to those disclosed herein. Modified nucleotides and modified internucleoside bonds

[046] The AAT RNAi agents disclosed herein may consist of modified nucleotides, which may preserve the activity of the RNAi agent while at the same time increasing serum stability and minimizing the possibility of activating interferon activity in humans. As used in this document, a “modified nucleotide” is a nucleotide other than a ribonucleotide (2'-hydroxyl nucleotide). In some embodiments, at least 50% (e.g., at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100%) of the nucleotides are modified nucleotides.As used in this document, modified nucleotides include any modified nucleotides known in the art, including, but not limited to, deoxyribonucleotides, nucleotide mimetics, 2'-modified nucleotides, inverted nucleotides, nucleotides comprising modified nucleobases, bridging nucleotides, nucleic acids. Petition 870260074731, dated 07 / 27 / 2026, page 27 / 97 16 / 46 peptides (PNAs), 2',3'-dry nucleotide mimetics (unlocked nucleobase analogs), locked nucleotides, 3'-O-methoxy nucleotides (2'-linked internucleosides), 2'-F-arabino nucleotides, 5'-Me, 2'-fluoro nucleotides, morpholino nucleotides, vinyl phosphonate-containing nucleotides, and cyclopropyl phosphonate-containing nucleotides. In some embodiments, the modified nucleotides of an AAT RNAi agent are 2'-modified nucleotides (i.e., a nucleotide with a group other than a hydroxyl group at the 2' position of the five-membered sugar ring). Modified 2'-nucleotides include, but are not limited to, 2'O-methyl nucleotides, 2'-deoxy-2'-fluoro nucleotides (commonly referred to simply as 2'-fluoro nucleotides), 2'-deoxy nucleotides, 2'-methoxyethyl nucleotide (2'-O-2-methoxyethyl), 2'-amino nucleotides, and 2'-alkyl nucleotides. Additional 2'-modified nucleotides are known in the art.It is not necessary for all nucleotides in a given RNAi agent to be uniformly modified. Furthermore, more than one modification can be incorporated into a single AAT RNAi agent or even into a single nucleotide of the same agent. The sense and antisense strands of the AAT RNAi agent can be synthesized and / or modified by methods known in the art. The modification in one nucleotide is independent of the modification in another nucleotide.

[047] In some embodiments, the nucleobase (often referred to simply as the “base”) can be modified. As is commonly used in the art, natural nucleobases include the primary purine bases adenine and guanine, and the primary pyrimidine bases cytosine, thymine, and uracil. A nucleobase can be modified to include, without limitation, universal bases, hydrophobic bases, promiscuous bases, expanded-size bases, and fluorinated bases. (See, for example, Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed. Wiley-VCH, 2008). The synthesis of such modified nucleobases (including phosphoramidite compounds that include modified nucleobases) is known in the art. Petition 870260074731, dated 07 / 27 / 2026, page 28 / 97 17 / 46

[048] Modified nucleobases include, for example, 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, (e.g., 2-aminopropyladenine, 5-propynyluracil or 5-propynylcytosine), 5-methylcytosine (5-me-C), 5-hydroxymethylcytosine, inosine, xanthine, hypoxanthine, 2-aminoadenine, 6-alkyl (e.g., 6-methyl, 6-ethyl, 6-isopropyl or 6-n-butyl) derivatives of adenine and guanine, 2-alkyl (e.g., 2-methyl, 2-ethyl, 2-isopropyl or 2-n-butyl) and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine, 2-thiocytosine, 5-halouracil, cytosine, 5-propynyl uracil, 5-propynylcytosine, 6-azo uracil, 6-azo cytosine, 6-azo thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-sulfhydryl, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo (e.g., 5-bromo), 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-aza-adenine, 7-deazaguanine, 7-deaza-adenine,3-deazaguanine and 3-deazaadenine.

[049] In some embodiments, all or substantially all nucleotides of an AAT RNAi agent are modified nucleotides. As used in this document, an RNAi agent in which substantially all nucleotides present are modified nucleotides is an RNAi agent with four or fewer (i.e., 0, 1, 2, 3, or 4) nucleotides in the sense and antisense strands being ribonucleotides (i.e., unmodified). As used in this document, a sense strand in which substantially all nucleotides present are modified nucleotides is a sense strand with two or fewer (i.e., 0, 1, or 2) nucleotides in the sense strand being ribonucleotides. As used in this document, a sense-antisense strand in which substantially all the nucleotides present are modified nucleotides is a sense-antisense strand having two or fewer (i.e., 0, 1, or 2) nucleotides in the sense strand being ribonucleotides.

[050] In some embodiments, one or more nucleotides of an AAT RNAi agent are linked by non-standard linkages or backbones (i.e., linkages Petition 870260074731, dated 07 / 27 / 2026, p. 29 / 97 18 / 46 modified internucleosides or modified backbones). Modified internucleosidic linkages or backbones include, but are not limited to, phosphorothioate groups, chiral phosphorothioates, thiophosphates, phosphorodithioates, phosphotriesters, aminoalkyl-phosphotriesters, alkyl phosphonates (e.g., methyl phosphonates or 3'-alkylene phosphonates), chiral phosphonates, phosphinates, phosphoramidates (e.g., 3'-amino phosphoramidate, aminoalkylphosphoramidates, or thionophosphoramidates), thionoalkyl-phosphonates, thionoalkylphosphotriesters, morpholino linkages, boranephosphates with normal 3'-5' linkages, 2'-5' linked boranephosphate analogs, or boranephosphates with inverted polarity, in which adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-3'. 5'-2'. In some embodiments, an internucleoside bond or modified backbone lacks a phosphorus atom.Modified internucleosidic linkages without a phosphorus atom include, but are not limited to, short-chain alkyl or cycloalkyl inter-sugar linkages, mixed heteroatoms and alkyl or cycloalkyl inter-sugar linkages, or one or more short-chain heteroatomic or heterocyclic inter-sugar linkages. In some embodiments, the modified internucleoside backbones include, but are not limited to, siloxane backbones, sulfide backbones, sulfoxide backbones, sulfone backbones, formacetyl and thioformacetyl backbones, methylene formacetyl and thioformacetyl backbones, alkene-containing backbones, sulfamate backbones, methyleneimino and methylenehydrazino backbones, sulfonate and sulfonamide backbones, amide backbones, and other backbones with N, O, S, and CH2 components.

[051] In some embodiments, a sense strand of an AAT RNAi agent may contain 1, 2, 3, 4, 5, or 6 phosphorothioate linkages, an antisense strand of an AAT RNAi agent may contain 1, 2, 3, 4, 5, or 6 phosphorothioate linkages, or both sense and antisense strands may independently contain 1, 2, 3, 4, 5, or 6 linkages Petition 870260074731, dated 07 / 27 / 2026, p. 30 / 97 19 / 46 phosphorothioate. In some embodiments, a sense strand of an AAT RNAi agent may contain 1, 2, 3, or 4 phosphorothioate linkages, an antisense strand of an AAT RNAi agent may contain 1, 2, 3, or 4 phosphorothioate linkages, or both sense and antisense strands may independently contain 1, 2, 3, or 4 phosphorothioate linkages.

[052] In some embodiments, an AAT RNAi agent sense strand contains at least two phosphorothioate internucleosidic bonds. In some embodiments, the at least two phosphorothioate internucleosidic bonds are between nucleotides at positions 1-3 of the 3' end of the sense strand. In some embodiments, the at least two phosphorothioate internucleosidic bonds are between nucleotides at positions 1-3, 2-4, 3-5, 4-6, 4-5, or 6-8 of the 5' end of the sense strand. In some embodiments, the phosphorothioate internucleosidic bonds are used to link terminal nucleotides in the sense strand to capping residues present at the 5' end, the 3' end, or both the 5' and 3' ends of the nucleotide sequence. In some embodiments, phosphorothioate internucleoside linkages are used to attach a target group to the sense strand.

[053] In some embodiments, an antisense strand of the AAT RNAi agent contains three or four phosphorothioate internucleosidic bonds. In some embodiments, an antisense strand of the AAT RNAi agent contains three phosphorothioate internucleosidic bonds. In some embodiments, the three phosphorothioate internucleosidic bonds are between nucleotides at positions 1-3 of the 5' end of the antisense strand and between nucleotides at positions 19-21, 20-22, 21-23, 22-24, 23-25, or 24-26 of the 5' end. In some embodiments, an AAT RNAi agent contains at least two phosphorothioate internucleosidic bonds in the sense strand and three or four phosphorothioate internucleosidic bonds in the antisense strand.

[054] In some embodiments, an AAT RNAi agent contains one or more Petition 870260074731, dated 07 / 27 / 2026, page 31 / 97 20 / 46 modified nucleotides and one or more modified internucleoside bonds. In some embodiments, a modified 2' nucleoside is combined with a modified internucleoside bond. Coating residues or fractions

[055] In some embodiments, the sense strand may include one or more capping residues or fractions, sometimes referred to in the art as a “cap,” a “terminal cap,” or a “capping residue.” As used in this document, a “capping residue” is a non-nucleotide compound or other fraction that may be incorporated into one or more terminals of a nucleotide sequence of an RNAi agent disclosed in this document. A capping residue may provide the RNAi agent, in some cases, with certain beneficial properties, such as, for example, protection against exonuclease degradation. In some embodiments, inverted abasic (invAb) residues (also referred to in the art as “inverted abasic sites”) are added as capping residues (see Table A). (See, for example, F. Czauderna, Nucleic Acids Res., 2003, 31(11), 270516).Capping residues are generally known in the art and include, for example, inverted abasic residues, as well as carbon chains such as C3H7 (propyl), C6H13 (hexyl), or C12H25 (dodecyl) groups. In some embodiments, a capping residue is present at the 5' terminal end, the 3' terminal end, or both the 5' and 3' terminal ends of the sense strand. In some embodiments, the 5' and / or the 3' end of the sense strand may include more than one inverted abasic deoxyribose moiety as a capping residue.

[056] In some embodiments, one or more inverted abasic residues (invAb) are added to the 3' end of the sense strand. In some embodiments, one or more inverted abasic residues (invAb) are added to the 5' end of the sense strand. In some embodiments, one or more inverted abasic residues or inverted abasic sites are inserted between the targeting ligand and the sequence of Petition 870260074731, dated 07 / 27 / 2026, page 32 / 97 21 / 46 nucleotides of the RNAi agent's sense strand. In some embodiments, the inclusion of one or more inverted abasic residues or inverted abasic sites at or near the terminal end or ends of the sense strand of an RNAi agent allows for enhanced activity or other desired properties of an RNAi agent.

[057] In some embodiments, one or more inverted abasic residues (invAb) are added to the 5' end of the sense strand. In some embodiments, one or more inverted abasic residues may be inserted between the targeting ligand and the nucleotide sequence of the RNAi agent's sense strand. The inverted abasic residues may be linked via phosphate, phosphorothioate (e.g., shown here as (invAb)s)), or other internucleosidic linkages. The chemical structures for inverted abasic deoxyribose residues are shown in Table A below, as well as in the chemical structures shown in Figures 1A to 1E and Figures 2A to 2E. Petition 870260074731, dated 07 / 27 / 2026, page 33 / 97 22 / 46

[059] An AAT RNAi agent may be conjugated to one or more non-nucleotide groups, including, but not limited to, a targeting fraction or a targeting group. A targeting fraction or targeting group may improve the targeting or distribution of the RNAi agent. Examples of targeting fractions and targeting groups are known in the art. Specific examples of the targeting group (NAG37)s used in the AAT RNAi Drug Substance described in Table 2 in this document, which includes three fractions Petition 870260074731, dated 07 / 27 / 2026, page 34 / 97 23 / 46 targeting groups of N-acetylgalactosamine are provided in Table B. The targeting moiety or targeting group may be covalently linked to the 3' and / or 5' end of the sense and / or antisense strand. In some embodiments, an AAT RNAi agent contains a targeting group attached to the 3' and / or 5' end of the sense strand. In some embodiments, a targeting group is attached to the 5' end of a sense strand of the AAT RNAi agent. In some embodiments, the targeting group comprises, consists essentially of, or consists of the (NAG37)s structure, and is attached to the 5' end of a sense strand of the AAT RNAi agent. A targeting group may be linked directly or indirectly to the RNAi agent via a ligand / linking group. In some embodiments, a targeting group is linked to the RNAi agent via a labile, cleavable, or reversible linker or ligand.In some embodiments, a targeting group is attached to an inverted abasic residue at the 5' end of the sense tape.

[060] Targeting groups or targeting fractions can enhance the pharmacokinetic or biodistribution properties of a conjugate or RNAi agent to which they are linked to improve cell-specific distribution and cell-specific uptake of the conjugate or RNAi agent. In some embodiments, a targeting group enhances the endocytosis of the RNAi agent. A targeting group can be monovalent, divalent, trivalent, tetravalent, or have a higher valence for the target to which it is directed. Representative targeting groups include, without limitation, compounds with affinity for cell surface molecules, cell receptor ligands, haptens, antibodies, monoclonal antibodies, antibody fragments, and antibody mimetics with affinity for cell surface molecules.

[061] In some embodiments, a targeting group comprises an asialoglycoprotein receptor ligand. In some embodiments, a ligand of Petition 870260074731, dated 07 / 27 / 2026, page 35 / 97 24 / 46 asialoglycoprotein receptor includes or consists of one or more galactose derivatives. As used in this document, the term galactose derivative includes galactose and galactose derivatives possessing affinity for the asialoglycoprotein receptor that is equal to or greater than that of galactose. Galactose derivatives include, but are not limited to: galactose, galactosamine, N-formylgalactosamine, N-acetylgalactosamine, N-propionylgalactosamine, Nn-butanoylgalactosamine, and Niso-butanoylgalactosamine (see, for example: ST Iobst and K. Drickamer, JBC, 1996, 271, 6686). Galactose derivatives and groups of galactose derivatives, which are useful for in vivo targeting of oligonucleotides and other molecules to the liver, are known in the art (see, for example, Baenziger and Fiete, 1980, Cell, 22, 611-620; Connolly et al., 1982, J. Biol. Chem., 257, 939-945).

[062] Galactose derivatives have been used to target molecules to hepatocytes in vivo through their binding to the asialoglycoprotein receptor expressed on the surface of hepatocytes. The binding of asialoglycoprotein receptor ligands to the asialoglycoprotein receptor(s) facilitates specific cell targeting to hepatocytes and endocytosis of the molecule into hepatocytes. Asialoglycoprotein receptor ligands can be monomeric (e.g., having a single galactose derivative) or multimeric (e.g., having multiple galactose derivatives). The galactose derivative or “cluster” of galactose derivative can be attached to the 3' or 5' end of the sense or antisense strand of the RNAi agent using methods known in the art.

[063] In some embodiments, a targeting group comprises a cluster of galactose derivatives. As used in this document, a cluster of galactose derivatives comprises a molecule with two to four terminal galactose derivatives. A terminal galactose derivative is linked to a molecule through its C-1 carbon. In some embodiments, the galactose derivative cluster is a galactose derivative trimer (also referred to as Petition 870260074731, dated 07 / 27 / 2026, p. 36 / 97 25 / 46 tri-antennary galactose derivative or trivalent galactose derivative). In some embodiments, the galactose-derived group comprises N-acetylgalactosamines. In some embodiments, the galactose-derived group comprises three N-acetylgalactosamines. In some embodiments, the galactose-derived group is a galactose-derived tetramer (also referred to as a tetra-antennary galactose derivative or tetravalent galactose derivative). In some embodiments, the galactose-derived group comprises four N-acetylgalactosamines.

[064] As used in this document, a galactose-derived trimer contains three galactose derivatives, each linked to a central branching point. As used in this document, a galactose-derived tetramer contains four galactose derivatives, each linked to a central branching point. The galactose derivatives may be linked to the central branching point through the C-1 carbons of the saccharides. In some embodiments, the galactose derivatives are linked to the branching point by means of linkers or spacers. In some embodiments, the linker or spacer is a flexible hydrophilic spacer, such as a PEG group (see, for example, US Patent 5,885,968; Biessen et al. J. Med. Chem. 1995 Vol. 39 p. 1538-1546). The branching point can be any small molecule that allows the binding of three galactose derivatives and still allows the branching point to bind to the RNAi agent.An example of a branch point group is di-lysine or di-glutamate. The linkage of the branch point to the RNAi agent can occur through a linker or spacer. In some embodiments, the linker or spacer comprises a flexible hydrophilic spacer, such as, but not limited to, a PEG spacer. In some embodiments, the linker comprises a rigid linker, such as a cyclic group. In some embodiments, a galactose derivative comprises or consists of N-acetylgalactosamine. In some embodiments, the group derived from... Petition 870260074731, dated 07 / 27 / 2026, p. 37 / 97 26 / 46 galactose is composed of a galactose-derived tetramer, which may be, for example, an N-acetylgalactosamine tetramer.

[065] The preparation of targeting groups, such as galactose derivative clusters including N-acetylgalactosamine, is described, for example, in International Patent Application Publication WO 2018 / 044350 (PCT / US2017 / 021147 Patent Application) and International Patent Application Publication WO 2017 / 156012 (PCT / US2017 / 021175 Patent Application), the contents of both of which are incorporated by reference herein in their entirety.

[066] For example, the targeting ligand conjugated with the AAT RNAi agent described in Tables 1A and 1B has the chemical structure of (NAG37)s, as shown in Table B. Petition 870260074731, dated 07 / 27 / 2026, p. 38 / 97 27 / 46

[068] In some embodiments, the AAT RNAi agent used in the methods disclosed in this document has the nucleotide sequences of the AAT RNAi Drug Substance (ADS-001) shown in Table 2. The nucleotide sequences of the AAT RNAi agent found in the AAT RNAi Drug Substance include an antisense strand nucleotide sequence as set forth in Table 1A below, and a sense strand nucleotide sequence as set forth in Table 1B follows.

[069] Table 1A. Antisense strand sequence of the AAT RNAi agent SEQ „ . . , ... . . |n Antisense sequence (modified) NO. <5· - 3') Base sequence ^S x«Q Underlying ID NO. (5' — 3') 2 usGfsuUfaAfacaugCfcUfaAfaCfgCf su UGUUAAACAUGCCUAAA 3 CGCU

[070] Table 1B. Nucleotide sequence of AAT RNAi agent detection chain (shown as a modified version without inverted abasic residues or NAG targeting group present in the AAT RNAi Drug Substance) SEQ ID NO. Sense Sequence (Modified) (5' — 3') SEQ ID NO. Underlying Basis Sequence (5' — 3') 4 agcguuuaGfGfCfauguuuaac 5 AGCGUUUAGGCAUGUU Petition 870260074731, dated 07 / 27 / 2026, p. 39 / 97 28 / 46 SEQ ID NO. Sense Sequence (Modified) (5' - 3') SEQ ID NO. Underlying Baseline Sequence (5' - 3') a UAACA

[071] As used in Tables 1A, 1B and 2 herein, the following notations are used to indicate modified nucleotides, targeting groups and linking groups: A, G, C and U represent adenosine, cytidine, guanosine or uridine; a, c, g and U represent 2'-O-methyl adenosine, cytidine, guanosine or uridine, respectively; Af, Cf, Gf and Uf represent 2'-fluoro adenosine, cytidine, guanosine or uridine, respectively; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic deoxyribose residue (see Table A); and (NAG37)s represents the structure shown in Table B above.

[072] As one skilled in the art would readily understand, unless otherwise indicated by the sequence (such as, for example, by a phosphorothioate linkage “s”), when present in a strand, the monomers are mutually linked by 5'-3'-phosphodiester linkages. As one skilled in the art would clearly understand, the inclusion of a phosphorothioate linkage, as shown in the modified nucleotide sequences disclosed herein, replaces the phosphodiester linkage typically present in oligonucleotides. Furthermore, one skilled in the art would readily understand that the terminal nucleotide at the 3' end of a given oligonucleotide sequence would typically have a hydroxyl group (-OH) at the respective 3' position of the given monomer instead of an ex vivo phosphate moiety.Furthermore, for the embodiments disclosed herein, when viewing the respective 5'^3' ribbon, the inverted abasic residues are inserted so that the 3' position of the deoxyribose is linked to the 3' end of the preceding monomer in the respective ribbon. Moreover, as one skilled in the art would readily understand and appreciate, while the phosphorothioate chemical structures represented herein typically show the anion on the sulfur atom, the inventions disclosed herein encompass all phosphorothioate tautomers (e.g., where the sulfur atom has a...). Petition 870260074731, dated 07 / 27 / 2026, page 40 / 97 29 / 46 double bond and the anion is on an oxygen atom). Unless expressly stated otherwise in this document, such understandings of those skilled in the art are used in describing the AAT RNAi agents and compositions that include AAT RNAi agents disclosed in this document.

[073] Each sense tape and / or antisense tape may have any of the above-listed direction groups or link groups, as well as other direction or link groups, conjugated to the 5' and / or 3' end of the sequence.

[074] The AAT agent RNAi antisense strand sequence is designed to target mRNA transcripts of normal and mutant AAT genes, thereby silencing the translation of mutant Z-AAT proteins using an RNA interference mechanism for human subjects with AATD.

[075] In some embodiments, the methods disclosed herein use the AAT RNAi Drug Substance established in the following Table 2:

[076] Table 2. Drug Substance RNAi of AAT (ADS-001)_____________ Sense and antisense strands (The sense and antisense strands are annealed to form a duplex): Sense strand (modified sequence) (5'^3'): Antisense strand (modified sequence) (5'^3'): (NAG37)s(invAb)sagcguuuaGfGfCfauguuuaacas(invAb) usGfsu UfaAfaca ug CfcUfaAfa Cfg Cfsu (SEQ ID NO:6) (SEQ ID NO:2) Chemical Formula: C493H610F11N163Na43O312P43S6 (Na+ form) C493H653F11N163O312P43S6 (H+ form) Molecular Weight: 16532.9 Da (Na+ form) 15587.6 Da (H+ form) Physical Appearance: White powder whitish.

[077] A schematic representation of the AAT RNAi Drug Substance (ADS-001) is shown in Figure 3, and complete representations of the chemical structure are shown in Figures 1A to 1E (sodium salt form) and Figures 2A to 2E (free acid form). In some embodiments, the AAT RNAi Drug Substance is prepared or supplied as a salt, mixed salt, or a free acid. In preferred embodiments, the form is a sodium salt. Petition 870260074731, dated 07 / 27 / 2026, page 41 / 97 30 / 46 Pharmaceutical Compositions and Formulations

[078] Suitable AAT RNAi agents for use in the methods disclosed herein may be prepared as pharmaceutical compositions or formulations for administration to human subjects. Pharmaceutical compositions may be used to treat a subject with a disease or disorder that would benefit from inhibition of AAT mRNA expression or reduction in AAT protein levels, such as human subjects with AATD. In some embodiments, the methods involve administering an AAT RNAi agent that is linked to a targeting group or targeting ligand, as described in this document, to a subject in need of treatment. In some embodiments, one or more pharmaceutically acceptable excipients (including vehicles, carriers, diluents, and / or delivery polymers) are added to pharmaceutical compositions that include an AAT RNAi agent, thus forming a pharmaceutical formulation suitable for in vivo delivery to a human subject.

[079] Pharmaceutical compositions that include an AAT RNAi agent, when administered to a human subject using the methods disclosed herein, decrease the level of AAT mRNA in the subject.

[080] In some embodiments, the described pharmaceutical compositions, including an AAT RNAi agent, are used to treat or manage clinical presentations in a subject with AATD, such as chronic hepatitis, cirrhosis, increased risk of hepatocellular carcinoma, transaminitis, cholestasis, fibrosis, and even fulminant hepatitis. In some embodiments, a therapeutically or prophylactically effective amount of one or more of the pharmaceutical compositions is administered to a subject in need of such treatment. In some embodiments, administration of any of the disclosed AAT RNAi agents may be used to decrease the number, severity, and / or frequency of symptoms of a disease in a subject. Petition 870260074731, dated 07 / 27 / 2026, page 42 / 97 31 / 46

[081] The described pharmaceutical compositions that include an AAT RNAi agent can be used to treat at least one symptom in a subject with a disease or disorder that would benefit from reduction or inhibition in AAT mRNA expression. In some embodiments, the subject is administered a therapeutically effective amount of one or more pharmaceutical compositions, including an AAT RNAi agent, thus treating the symptom. In other embodiments, the subject is administered a prophylactically effective amount of one or more AAT RNAi agents, thus preventing at least one symptom.

[082] The AAT RNAi agents disclosed in this document can be administered via any suitable route in a preparation appropriately adapted to the particular route. Thus, the pharmaceutical compositions described herein can be administered by injection, for example, intravenously or subcutaneously. In some embodiments, the pharmaceutical compositions described herein are administered by subcutaneous injection.

[083] As used in this document, a pharmaceutical composition or drug product includes a pharmacologically effective amount of at least one AAT RNAi agent and one or more pharmaceutically acceptable excipients. Pharmaceutically acceptable excipients (excipients) are substances other than the Active Pharmaceutical Ingredient (API, therapeutic product, e.g., AAT RNAi agent) that are intentionally included in the drug delivery system. Excipients do not exert or are not intended to exert a therapeutic effect at the intended dosage. Excipients may act to a) assist in the processing of the drug delivery system during manufacture, b) protect, support or enhance the stability, bioavailability or patient acceptability of the API, c) assist in product identification and / or d) enhance any other attribute of the overall safety, delivery efficacy of the API during storage or use.A pharmaceutically acceptable excipient may or may not be an inert substance. Petition 870260074731, dated 07 / 27 / 2026, page 43 / 97 32 / 46

[084] Excipients may include, but are not limited to: absorption enhancers, anti-adherents, anti-foaming agents, antioxidants, binders, buffering agents, carriers, coating agents, colors, distribution enhancers, distribution polymers, dextran, dextrose, diluents, disintegrants, emulsifiers, extenders, fillers, flavors, glidants, humectants, lubricants, oils, polymers, preservatives, physiological saline, salts, solvents, sugars, suspending agents, sustained-release matrices, sweeteners, thickening agents, tonicity agents, vehicles, water-repellent agents and humectants.

[085] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (when soluble in water). For subcutaneous or intravenous administration, suitable carriers may include physiological saline, bacteriostatic water, Cremophor® EL™ (BASF, Parsippany, NJ) or phosphate-buffered saline (PBS). It should be stable under manufacturing and storage conditions and should be preserved against contaminating action by microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), and appropriate mixtures thereof.

[086] Sterile injectable solutions can be prepared by incorporating the active compound in the required quantity into an appropriate solvent with one or a combination of ingredients listed above, as required, followed by sterilization by filtration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and the other required ingredients from those listed above.

[087] In some embodiments, a pharmaceutical composition suitable for use in the methods disclosed in this document includes the components Petition 870260074731, dated 07 / 27 / 2026, page 44 / 97 33 / 46 identified in the formulated AAT RNAi Drug Substance provided in Table 3 below.

[088] RNAi AAT agents may be formulated in unit dosage compositions for ease of administration and dosage uniformity. The unit dosage form refers to physically distinct units suitable as unit dosages for the subject to be treated; each unit contains a predetermined amount of the therapeutic compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. In some embodiments, the unit dosage is between about 5 mg and about 300 mg of AAT RNAi Drug Substance. In some embodiments, the unit dosage is between about 25 mg and about 200 mg of AAT RNAi Drug Substance. In some embodiments, the unit dosage is between about 100 mg and about 200 mg of AAT RNAi Drug Substance. In some embodiments, the unit dosage is about 100 mg of AAT RNAi Drug Substance.In some formulations, the dosage unit is approximately 200 mg of AAT RNAi Drug Substance.

[089] A pharmaceutical composition may contain other additional components commonly found in pharmaceutical compositions. These additional components include, but are not limited to: antipruritic agents, astringents, local anesthetics or anti-inflammatory agents (e.g., antihistamine, diphenhydramine, etc.).

[090] As used in this document, “pharmacologically effective amount”, “therapeutically effective amount” or simply “effective amount” refers to the amount of an RNAi agent to produce a pharmacological, therapeutic or preventive result.

[091] The pharmaceutically acceptable formulations described may be packaged in kits, containers, packages or dispensers. The compositions Petition 870260074731, dated 07 / 27 / 2026, page 45 / 97 34 / 46 pharmaceuticals described here can be packaged in pre-filled syringes or vials. AAT-formulated RNAi drug substance

[092] In some embodiments, the AAT RNAi Drug Substance as provided in Table 2 (ADS-001) is formulated with one or more pharmaceutically acceptable excipients to form a pharmaceutical composition suitable for administration to a human subject. In some embodiments, the AAT RNAi Drug Substance described in Table 2 is formulated at 230 mg / mL in an aqueous sodium phosphate buffer (0.5 mM monobasic sodium phosphate, 0.5 mM dibasic sodium phosphate), forming the formulated AAT RNAi Drug Substance (ADS-0011) shown in Table 3:

[093] Table 3. Composition of the formulated AAT RNAi drug substance, per 1.0 mL Component Function Quality / Grade Concentration ADS-001 Active ingredient Internal 230 mg Monobasic sodium phosphate, monohydrate Suspending agent USP, Ph. Eur 0.061 mg Dibasic sodium phosphate, anhydrous Suspending agent USP, Ph. Eur 0.062 mg Water for injection (WFI) Vehicle USP, Ph. Eur 879.2 mg

[094] The AAT RNAi Drug Substance formulated according to Table 3 is prepared as a sterile formulation. In some embodiments, the formulated AAT RNAi Drug Substance is packaged in a container, such as a glass vial. In some embodiments, the formulated AAT RNAi Drug Substance is packaged in a glass vial with a fill volume of approximately 1.1 mL, and a desired volume for administration can be calculated based on the desired dose level to be administered.

[095] In some embodiments, the formulated AAT RNAi Drug Substance presented in Table 3 is administered to a human subject using the Petition 870260074731, dated 07 / 27 / 2026, p. 46 / 97 35 / 46 methods disclosed here. Human subjects diagnosed with AATD and AATD

[096] The methods disclosed in this document include treating alpha-1 antitrypsin deficiency (AATD) in a human subject in need thereof, including treatment of symptoms and diseases caused by AATD in the human subject, using pharmaceutical compositions that include the AAT RNAi Drug Substance described in Table 2. In some embodiments, the human subject is diagnosed with AATD prior to administration. As noted herein, AATD is a genetic disorder caused by mutations in the gene transcript that result in the translation of a mutant form of the AAT protein, for which some mutant forms that are prone to abnormal folding lead to intracellular retention in hepatocytes. Although several mutations of the SERPINA1 gene have been identified, the most common and serious form of AATD, the PiZZ genotype, is caused by a single base pair substitution.In subjects with the PiZZ genotype, circulating AAT levels are frequently reported as less than 15% of levels in normal humans. In many cases, patients are initially diagnosed with COPD, asthma, or another lung disease without identification of the underlying cause. Over time, liver diseases such as fibrosis and cirrhosis may develop due to intercellular retention of the misfolded protein (“Z-AAT”) and the inability to adequately secrete the protein from liver cells. Pediatric patients often present with clinical symptoms of liver disease, which may include asymptomatic chronic hepatitis, failure to thrive, poor nutrition, or hepatomegaly and splenomegaly. AATD can be diagnosed and confirmed through standard genotyping of blood samples from the subject. Dosage and inhibition of AAT gene expression

[097] Generally, an effective amount of an AAT RNAi agent will be in the range of about 0.1 to about 10 mg / kg of body weight / day, for example, of Petition 870260074731, dated 07 / 27 / 2026, page 47 / 97 36 / 46 approximately 0.25 to approximately 5 mg / kg of body weight / day. In some embodiments, an effective amount of an AAT RNAi agent will be in the range of approximately 0.5 to approximately 4 mg / kg of body weight per dose. In some embodiments, the effective amount is a fixed dose. In some embodiments, a fixed dose of 5 mg to 300 mg of AAT RNAi Drug Substance is an effective dose. In some embodiments, a fixed dose of 25 mg to 200 mg of AAT RNAi Drug Substance is an effective dose. The amount administered will likely depend on variables such as the patient's general age and health status, the relative biological efficacy of the compound administered, the drug formulation, the presence and types of excipients in the formulation, and the route of administration.In some forms, a fixed dose of approximately 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 120, 140, 160, 180, 200, 220, 240, 260 or 280 mg is given. 260, 280, or 300 mg is an effective dose. In some embodiments, a fixed dose of approximately 25 mg, approximately 100 mg, or approximately 200 mg is an effective dose.

[098] Furthermore, it should be understood that the initial dosage administered may, in some cases, be increased beyond the upper level above to rapidly achieve the desired blood or tissue level, or the initial dosage may, in some cases, be lower than ideal. For example, in some embodiments, an initial dose of about 25 mg to about 200 mg of AAT RNAi Drug Substance is administered, followed by a second dose of about 25 to 200 mg of AAT RNAi Drug Substance approximately 1 month later, and thereafter, additional doses (a concept similar to “maintenance doses”) are administered once every three months (i.e., once per quarter).

[099] For the treatment of the disease or for the formation of a medicament or composition for the treatment of a disease, the pharmaceutical compositions described herein, including an AAT RNAi agent, may be combined with a Petition 870260074731, dated 07 / 27 / 2026, p. 48 / 97 37 / 46 excipient or with a second therapeutic agent or treatment including, but not limited to: a second or other RNAi agent, a small molecule drug, an antibody, an antibody fragment, peptide and / or aptamer.

[0100] In some embodiments, the gene expression level and / or mRNA level of an AAT gene in a subject to whom a described AAT RNAi agent is administered is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, 99%, or more than 99% relative to the subject before the AAT RNAi agent is administered or to a subject who has not received the AAT RNAi agent. The gene expression level and / or mRNA level in the subject is reduced in a cell, group of cells, and / or tissue of the subject.

[0101] In some embodiments, the AAT protein level in a subject to whom a described AAT RNAi agent has been administered is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or greater than 99% relative to the subject before the AAT RNAi agent was administered or to a subject who did not receive the AAT RNAi agent. The protein level in the subject is reduced in a cell, group of cells, tissue, blood and / or other fluid of the subject.

[0102] In some embodiments, the protein level of the Z-AAT polymer in a subject with AATD who has been administered a described AAT RNAi agent is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more than 99% compared to the subject before the AAT RNAi agent was administered or to a subject who did not receive the AAT RNAi agent. In some embodiments, the protein level of the Z-AAT polymer in a subject to whom a described AAT RNAi agent has been administered is reduced by at least approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, Petition 870260074731, dated 07 / 27 / 2026, p. 49 / 97 38 / 46 96%, 97%, 98%, 99% or greater than 99% compared to the subject before the AAT RNAi agent was administered or to a subject who did not receive the AAT RNAi agent.

[0103] A reduction in AAT gene expression, AAT mRNA, or AAT protein levels can be assessed and quantified by general methods known in the art. The examples disclosed here present generally known methods for assessing the inhibition of AAT gene expression and the reduction of AAT protein levels. The reduction or decrease in AAT mRNA level and / or protein level (including Z-AAT polymer and / or monomer) are collectively referred to in this document as a reduction or decrease in AAT or inhibition or reduction of AAT expression.

[0104] As used in this document, the terms “treat,” “treatment,” and the like mean the methods or steps taken to provide relief or reduction of the number, severity, and / or frequency of one or more symptoms of a disease in a subject. As used in this document, “treat” and “treatment” may include the prevention, management, prophylactic treatment, and / or inhibition of the number, severity, and / or frequency of one or more symptoms of a disease in a subject.

[0105] As used in this document, “monthly dosage” or “monthly” administration means every 28 days. As used in this document, “quarterly dosage” or “quarterly” administration means every 84 days. The term “approximately” when used in connection with monthly dosage means monthly dosage + / - 3 days. The term “approximately” when used in connection with quarterly dosage means quarterly dosage + / - 9 days.

[0106] As used in this document, the phrase “introduction into a cell,” when referring to an RNAi agent, means functionally distributing the RNAi agent into a cell. The phrase “functional distribution” means distributing the RNAi agent to the cell in a way that allows the RNAi agent to have the activity Petition 870260074731, dated 07 / 27 / 2026, page 50 / 97 39 / 46 expected biological effect, for example, sequence-specific inhibition of gene expression.

[0107] Unless otherwise indicated, the use of the symbol as used in this document means that any group or groups may be linked to the same that is consistent with the scope of the inventions described herein.

[0108] As used in this document, unless specifically identified in a structure as having a particular conformation, for each structure in which asymmetric centers are present and thus give rise to enantiomers, diastereomers, or other stereoisomeric configurations, each structure disclosed here is intended to represent all possible isomers, including their optically pure and racemic forms. For example, the structures disclosed here are intended to cover mixtures of diastereomers as well as individual stereoisomers.

[0109] As used in a claim herein, the phrase “consisting of” excludes any element, step, or ingredient not specified in the claim. When used in a claim herein, the phrase “essentially consisting of” limits the scope of a claim to the materials or steps specified and those that do not materially affect the basic and novel features of the claimed invention.

[0110] Those skilled in the art would readily understand and appreciate that the compounds and compositions disclosed herein may have certain atoms (e.g., N, O, or S atoms) in a protonated or deprotonated state, depending on the environment in which the compound or composition is placed. Consequently, as used in this document, the structures disclosed herein consider that certain functional groups, such as, for example, OH, SH, or NH, may be protonated or deprotonated. The disclosure in this document is intended to cover the disclosed compounds and compositions regardless of their protonation state based on Petition 870260074731, dated 07 / 27 / 2026, p. 51 / 97 40 / 46 in the environment (such as pH), as would be easily understood by someone versed in the technique.

[0111] Unless otherwise defined, all technical and scientific terms used in this document have the same meaning as commonly understood by one skilled in the art. Although methods and materials similar or equivalent to those described in this document may be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned in this document are incorporated in their entirety by reference. In case of conflict, this descriptive report, including definitions, shall serve as the basis for control. Furthermore, the materials, methods, and examples are for illustrative purposes only and are not intended to be limiting.

[0112] The modalities and items provided above are now illustrated with the following non-limiting examples. Examples Example 1. Synthesis and Formulation of the RNAi Drug Substance from AAT (ADS-001)

[0113] The AAT RNAi Drug Substance suitable for use in the methods disclosed herein can be synthesized using standard phosphoramidite technology in solid-phase oligonucleotide synthesis as is known in the art. Commercially available oligonucleotide synthesizers (e.g., MerMade96E® (Bioautomation) or MerMade12® (Bioautomation)) can be used. Syntheses can be performed on a solid support made of controlled pore glass (CPG, 500 Å or 600 Å, obtained from Prime Synthesis, Aston, PA, USA). The monomer positioned at the 3' end of the respective strand can be fixed to the solid support as a starting point for synthesis. All RNA, 2' modified RNA phosphoramidites, and inverted abasic phosphoramidites can be acquired Petition 870260074731, dated 07 / 27 / 2026, page 52 / 97 41 / 46 commercially. Phosphoramides containing a targeting group can be synthesized that are suitable for addition to the 5' end of the sense strand. Standard cleavage, deprotection, purification, and annealing steps can be used as known in the art. Further description relating to the synthesis of AAT RNAi agents can be found, for example, in International Patent Application Publication WO 2018 / 132432 (PCT / US2018 / 013102) and WO 2018 / 044350 (PCT / US2017 / 021147), each of which is incorporated by reference herein in its entirety. The AAT RNAi Drug Substance can then be formulated by dissolving in standard pharmaceutically acceptable excipients that are generally known in the art. For example, Table 3 shows a formulated AAT RNAi Drug Substance that is suitable for use in the methods disclosed herein. Example 2. Phase I Clinical Trial of the AAT RNAi Drug Substance (ADS-001) in normal healthy human volunteers (NHV).

[0114] A Phase 1, single-dose, multiple-dose escalation study was conducted to evaluate the safety, tolerability, pharmacokinetics, and effect of the AAT RNAi Drug Substance (ADS-001) on serum AAT levels in healthy volunteers (NHV). The study subject population included healthy adult men and women aged 18–52 years with BMI between 19.0 and 35.0 kg / m2.

[0115] Subjects with NHV were divided into a total of seven cohorts. Cohorts 1 to 4 were randomized to receive AAT RNAi Drug Substance or placebo (4 active: 4 placebo) in single increasing doses of 35 mg (Cohort 1) and multiple increasing doses of 100 mg (Cohort 2), 200 mg (Cohort 3), and 300 mg (Cohort 4) administered as a subcutaneous injection. Cohorts 1 to 4 were double-blind. Cohorts 2b, 3b, and 4b were open-label, consisting of 4 subjects who received single doses of 100, 200, and 300 mg of AAT RNAi Drug Substance. A total of 44 subjects completed the study. Figure 4 shows the final study design. Petition 870260074731, dated 07 / 27 / 2026, page 53 / 97 42 / 46 for the Phase I Clinical Trial. The study parameters are summarized in Table 4 below.

[0116] Table 4. Phase I Clinical Study Parameters Development Phase Phase 1: First in Human Study Objectives Primary Objectives: • To determine the incidence and frequency of adverse events possibly or probably related to treatment as a measure of the safety and tolerability of the AAT RNAi Drug Substance (ADS-001) using single-dose and multiple-dose escalating doses in normal healthy human volunteers (NHV). Secondary Objectives: • To evaluate the single-dose and multiple-dose pharmacokinetics of the AAT RNAi Drug Substance in NHV. • To determine the reduction in serum AAT in response to the AAT RNAi Drug Substance as a measure of drug activity.Exploratory Objectives: • To evaluate the effect of single doses of AAT RNAi Drug Substance on cytokines (cytokine panel A: interleukin-6 [IL-6], monocyte chemoattractant protein-1 [MCP-1], tumor necrosis factor alpha [TNF-alpha], interleukin-8 [IL-8], interleukin-1beta [IL1beta], interferon alpha [IFN alpha], IL-10, IL-12 [p40], IL-12 [p70], macrophage inflammatory protein-1alpha [Mip1alpha]) in NHV. • To evaluate the effect of increasing single doses of AAT RNAi Drug Substance on complement factors Bb, CH50, C5a, C4a, and C3a in NHV. • To collect plasma samples in NHV for subsequent metabolite identification (reported in a separate report outside of this study). • Collect urine samples at the NHV for subsequent determination of urinary excretion and metabolite identification (reported in a separate report outside of this study).Study design Cohorts 1 to 4: randomized, double-blind, placebo-controlled Cohorts 2b, 3b and 4b: open-label Study population This study was conducted in NHVs, adult males and females, aged 18 to 52 years and with a BMI between 19.0 and 35.0 kg / m2. AAT RNAi Drug Substance Product (ADS-001) (see. Petition 870260074731, dated 07 / 27 / 2026, page 54 / 97 43 / 46 Investigation Table 2), administered as formulated AAT RNAi Drug Substance (see Table 3) Dosage and frequency Cohort 1: randomized to receive AAT RNAi Drug Substance (ADS-001) or placebo (4 active: 4 placebo) in a single 35 mg dose administered as a single subcutaneous injection. Cohorts 2-4: randomized to receive three monthly doses (i.e., days 1, 29, and 57) of 100 mg (Cohort 2), 200 mg (Cohort 3), or 300 mg (Cohort 4) of AAT RNAi Drug Substance or placebo (4 active: 4 placebo), via subcutaneous injection. Cohorts 2b, 3b and 4b: enrolled to receive a single dose of 100 mg (Cohort 2b), 200 mg (Cohort 3b) or 300 mg (Cohort 4b) of AAT RNAi Drug Substance (4 actives) administered as a single subcutaneous injection. Placebo Reference Formulation (PBO): normal saline solution (0.9%) administered subcutaneously with a corresponding volume.Safety Assessment Criteria • Safety was assessed by adverse events, serious adverse events, physical examinations, vital sign measurements (blood pressure, heart rate, temperature, and respiratory rate), resting ECG measurements, clinical laboratory tests, concomitant medications / therapy, injection site reactions (ISRs), reasons for treatment discontinuation, and pregnancy follow-up 90 days after Day 29 (Cohort 1) and after Day 113 (all other cohorts). Pharmacokinetic Assessment Blood samples will be collected from each subject for pharmacokinetic analysis after dose 1 (Cohort 1) and after doses 1 and 3 (Cohorts 2, 3, and 4). Data Analysis Screening, compliance, tolerability, and safety data: Safety analyses will be performed and results summarized by cohort.The incidence and frequency of adverse events (AEs), serious adverse events (SAEs), related AEs, related SAEs, and AEs leading to discontinuation will be summarized by cohort by SOC, PT, and severity. Other safety parameters will be summarized at each scheduled time. Pharmacokinetics (NHV subjects only): Plasma concentrations of the constituents of the RNAi drug substance of AAT will be used for... Petition 870260074731, dated 07 / 27 / 2026, page 55 / 97 44 / 46 Calculate the following PK parameters: observed maximum plasma concentration (Cmax), area under the plasma concentration-time curve (AUC) from time 0 to 24 hours (AUC0-24), AUC from time 0 extrapolated to infinity (AUCinf), and terminal elimination half-life (t½). Pharmacokinetic parameters will be determined using non-compartmental methods. Descriptive statistics of PK parameters will include mean, standard deviation (SD), coefficient of variation, median, minimum, and maximum. PK results will be analyzed for dose proportionality and sex differences.

[0117] The serum AAT reduction results from the study showed that administration of the AAT RNAi Drug Substance at doses of 35 to 300 mg resulted in a profound reduction in serum AAT compared to placebo. Initially, a cohort was proposed as part of the clinical trial protocol of 400 mg of AAT RNAi Drug Substance per dose. However, in view of the unexpected potency at doses of 35, 100, 200, and 300 mg, the 400 mg cohort was removed from the study protocol. Doses of 35 mg, 100 mg, and 200 mg produced substantial reductions in serum AAT, with 100 mg and 200 mg achieving a mean serum AAT reduction of approximately 90% after multiple doses in the Phase I study. Figures 5 to 11 report the serum AAT reductions of the various cohorts in the Phase I study.

[0118] There was no clear dose-dependent response at all dose levels because, surprisingly and unexpectedly, the 100 mg and 200 mg dose levels produced substantial knockdown (reaching approximately 90%) and similar knockdown for the highest dose of 300 mg. Although the lowest dose of 35 mg was still quite active, it was not as active as 100 mg administered as a single dose, indicating a degree of dose response.

[0119] The duration of the reduction in serum AAT (> 58%) from a single 35 mg dose lasted longer than initially predicted, up to 16 weeks after dose administration with subsequent return to baseline. For example, thirty-four weeks after the single 35 mg dose, a subject's serum AAT level Petition 870260074731, dated 07 / 27 / 2026, p. 56 / 97 Subject 45 / 46's AAT level returned above 90 mg / dL, while the second subject's AAT level remained at 40 mg / dL (60.4% reduced from baseline). There was no significant difference in the duration of response from single doses of 100 mg to 300 mg of AAT RNAi Drug Substance, with return to baseline beginning between 8 and 16 weeks after the single dose.

[0120] Multiple doses of AAT RNAi Drug Substance maintain the profound reduction in serum AAT for a longer period than a single dose generally. These data suggest that a second dose received on Day 29 (i.e., one month after an initial dose) may further reduce serum AAT levels or maintain reductions, and subsequent doses may be administered to maintain maximum serum AAT reduction every 12 weeks (i.e., quarterly).

[0121] In the Phase I study, there were no deaths, no serious adverse events (SAEs), and no adverse events (AEs) classified as severe in intensity. Two subjects reported three AEs of moderate intensity in subjects receiving the AAT RNAi Drug Substance (upper respiratory tract infection, rhinorrhea, general chest pain). Three subjects reported three AEs as moderate in intensity in subjects receiving placebo (2-gastroenteritis, left-sided musculoskeletal chest pain). All other AEs were reported as mild. Most subjects reported AEs unrelated to the study treatment. One AE occurred in a subject receiving AAT, leading to premature discontinuation of therapy, although the subject continued to be followed in the study. Ninety-four AEs were reported in 28 subjects who received at least a single dose of the formulated AAT RNAi Drug Substance. Forty-six adverse events (AEs) were reported in 17 subjects who received placebo.There is no clear pattern of increased frequency or intensity of AEDs with increasing dose.

[0122] Six injection site AEs occurred in 6 subjects across all AAT RNAi Drug Substance cohorts, all occurring in Petition 870260074731, dated 07 / 27 / 2026, page 57 / 97 46 / 46 subjects received the drug. There were no injection site AEs in placebo subjects. Reported injection site reactions included bruising, erythema, and pain. These combined injection site AEs were reported by 21.4% of subjects who received the formulated AAT RNAi Drug Substance. Six of the 50 injections of the formulated AAT RNAi Drug Substance resulted in an injection site AE, or 12%. No injection site AEs were reported more than once in a single subject. All injection site AEs were considered mild in intensity. Other Modalities

[0123] It should be understood that, although the invention has been described in conjunction with its detailed description, the preceding description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, other advantages and other modifications are within the scope of the following claims. Petition 870260074731, dated 07 / 27 / 2026, p. 58 / 97

Claims

1 / 4 CLAIMS 1. Use of a pharmaceutical composition comprising the AAT RNAi Drug Substance described in Table 2, CHARACTERIZED in that it is in the manufacture of a medicament for treating alpha-1 antitrypsin deficiency (AATD) in a human individual in need thereof, wherein the pharmaceutical composition is formulated to be administered at a dose between about 5 mg and about 300 mg of the AAT RNAi Drug Substance, and wherein the medicament is formulated for administration once a month by subcutaneous injection.

2. Use of a pharmaceutical composition comprising the AAT RNAi Drug Substance described in Table 2, CHARACTERIZED in that it is in the manufacture of a medicament for treating alpha-1 antitrypsin deficiency (AATD) in a human individual in need thereof, wherein the pharmaceutical composition is formulated to be administered at a dose between about 5 mg and about 300 mg of the AAT RNAi Drug Substance, and wherein the medicament is formulated for administration once every three months by subcutaneous injection.

3. Use of a pharmaceutical composition comprising the AAT RNAi Drug Substance described in Table 2, CHARACTERIZED in that it is in the manufacture of a medicament to treat alpha-1 antitrypsin deficiency (AATD) in a human individual in need thereof, wherein the use comprises: a. formulating for administration to the individual subcutaneously of an initial dose of the pharmaceutical composition comprising the AAT RNAi Drug Substance at a dose of between about 5 mg and about 300 mg of the AAT RNAi Drug Substance, b. formulating for administration to the individual of a second dose of the pharmaceutical composition approximately one month after the initial dose, and c. formulating for administration to the individual of a third dose of the pharmaceutical composition approximately three months after the second dose. Petition 870260074731, dated 07 / 27 / 2026, page 59 / 97 2 / 4 4. Use, according to any of claims 1 to 3, CHARACTERIZED by the fact that the dose of the RNAi Drug Substance of AAT is between about 25 mg and about 300 mg.

5. Use, according to any of claims 1 to 3, CHARACTERIZED by the fact that the dose of the RNAi Drug Substance of AAT is between about 25 mg and about 200 mg.

6. Use, according to any of claims 1 to 3, CHARACTERIZED by the fact that the dose of the RNAi Drug Substance of AAT is between about 100 mg to about 200 mg.

7. Use, according to any of claims 1 to 3, CHARACTERIZED by the fact that the dose of the RNAi Drug Substance of AAT is between approximately 25 mg.

8. Use, according to any of claims 1 to 3, CHARACTERIZED by the fact that the dose of the RNAi Drug Substance of AAT is approximately 35 mg.

9. Use, according to any of claims 1 to 3, CHARACTERIZED by the fact that the dose of the RNAi Drug Substance of AAT is between approximately 100 mg.

10. Use, according to any of claims 1 to 3, CHARACTERIZED in that the dose of the RNAi Drug Substance of AAT is approximately 200 mg.

11. Use, according to any of claims 1 to 3, CHARACTERIZED in that the dose of the RNAi Drug Substance of AAT is between about 200 mg or less.

12. Use, according to any of claims 1 to 11, CHARACTERIZED by the fact that the condition or disease caused by AATD is a liver disease. Petition 870260074731, dated 07 / 27 / 2026, page 60 / 97 3 / 4 13. Use, according to claim 12, CHARACTERIZED by the fact that the liver disease is chronic hepatitis, cirrhosis, increased risk of hepatocellular carcinoma, transaminitis, cholestasis, fibrosis or fulminant hepatic failure.

14. Use, according to any one of claims 1 to 13, CHARACTERIZED in that an additional therapeutic for the treatment of AATD is formulated to be administered to the individual.

15. Use, according to any one of claims 1 to 14, CHARACTERIZED in that a therapeutic is further formulated to be administered to an individual for the treatment of lung damage, emphysema, or other lung diseases or disorders caused by deficiency of endogenously secreted AAT protein.

16. Use according to claim 15, CHARACTERIZED in that the additional therapeutic comprises human AAT protein, purified human alpha-1 proteinase inhibitor or recombinant AAT protein.

17. Use, according to any one of claims 1 to 16, CHARACTERIZED in that the pharmaceutical composition is packaged in a kit, container, packaging, dispenser, pre-filled syringe or vials.

18. Use, according to any of claims 1 to 17, CHARACTERIZED in that the pharmaceutical composition comprises, consists of, or essentially consists of the formulated AAT RNAi Drug Substance described in Table 3.

19. Use according to claim 3, CHARACTERIZED in that it further comprises formulating additional doses to be administered after the third dose, wherein the additional doses are formulated to be administered approximately three months apart.

20. Use, according to any of claims 1 to 19, CHARACTERIZED by the fact that the administration of one or more doses of the pharmaceutical composition is carried out by the individual.

21. Use, according to any of claims 1 to 19, CHARACTERIZED in that the administration of one or more doses of the pharmaceutical composition is carried out by a medical professional.

22. Use of the AAT RNAi Drug Substance described in Table 2 or of a pharmaceutical composition comprising the AAT RNAi Drug Substance described in Table 2, CHARACTERIZED in that it is in the manufacture of a medicament for treating alpha-1 antitrypsin deficiency (AATD) in a human individual in need thereof, wherein the use comprises formulating the pharmaceutical composition to be administered to the individual at a dose between about 5 mg and about 300 mg of the AAT RNAi Drug Substance, wherein the medicament is formulated to be administered once a month by subcutaneous injection.

23. Use of the AAT RNAi Drug Substance described in Table 2 or of a pharmaceutical composition comprising the AAT RNAi Drug Substance described in Table 2, CHARACTERIZED in that it is in the manufacture of a medicament for treating alpha-1 antitrypsin deficiency (AATD) in a human individual in need thereof, wherein the use comprises formulating the pharmaceutical composition to be administered to the individual at a dose between about 5 mg and about 300 mg of the AAT RNAi Drug Substance, wherein the medicament is formulated to be administered once every three months by subcutaneous injection.

24. Invention of a product, process, system, kit or use, CHARACTERIZED by the fact that it has one or more elements disclosed in the patent application.

25. Pharmaceutical composition CHARACTERIZED by the fact that it comprises the RNAi drug substance of AAT and one or more pharmaceutically acceptable excipients. Petition 870260074731, dated 07 / 27 / 2026, page 62 / 97

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