Compounds and adjuvant formulations useful in pneumococcal vaccines

Abstract

The present invention relates to novel compounds and formulations. Additionally, the present invention relates to novel compounds and formulations useful for pneumococcal and pneumococcal conjugate vaccines. More specifically, the present invention relates to compositions comprising a pneumococcal conjugate and one or more compounds of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or pharmaceutically acceptable salts thereof, formulated as a stable nanoemulsion (referred to herein as an "SNE adjuvant composition" or "SNE").

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 503,021, filed May 18, 2023, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] Pneumococcal disease is caused by infection with the bacterium Streptococcus pneumoniae (pneumococcus). Different pneumococcal serotypes are known to cause different manifestations of disease, and infections can result in a range of symptoms, from ear and sinus infections to pneumonia and bloodstream infections. Pneumococcal disease has high associated morbidity and mortality rates worldwide, particularly among the elderly and children. Currently, 100 capsular polysaccharides, some of which are used to define or designate specific serotypes, have been identified (Ganaie, F. et al. (2020) Clinical Science and Epidemiology, Vol. 11, Issue 3, pages 1-15). These serotypes are distinguished by their chemical structure, serological response, and other associated genetic variations.

[0003] In 1983, PNEUMOVAX® (Merck & Co., Inc., Rahway, New Jersey, USA), a 23-valent pneumococcal vaccine (PV), was approved in the United States. This vaccine demonstrated reduced immunogenicity in infants due to a T-cell-independent response. To address this issue, pneumococcal conjugate vaccines (PCVs) were developed, particularly in infants. By covalently linking polysaccharides to carrier proteins, the immunogenic response became T-cell dependent. In 2000, PREVNAR® (Pfizer Inc., Philadelphia, Pennsylvania, USA), a 7-valent PCV, was approved in the United States. In 2010, PREVNAR13® (Pfizer Inc.), a 13-valent PCV, was approved in the United States. In 2021, VAXNEUVANCE® (Merck & Co., Inc., Rahway, New Jersey, USA), a 15-valent PCV, and PREVNAR20® (Pfizer Inc.), a 20-valent PCV, were approved in the United States. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Ganaie,F.et al.(2020)Clinical Science and Epidemiology,Vol.11,Issue 3,pages 1-15 Summary of the Invention [Problem to be solved by the invention]

[0005] Licensed PCVs currently utilize aluminum-containing adjuvants to enhance immunogenicity. While aluminum adjuvants increase the immunogenic response from baseline, it is unclear whether the immunogenic response is sufficient for higher-valency PCVs, especially in infants. Therefore, there is a need to identify other adjuvants that can increase the immunogenicity of multivalent PCVs beyond the current aluminum adjuvant standard. [Means for solving the problem]

[0006] The present invention relates to formulations and compositions comprising (i) one or more Streptococcus pneumoniae (S. pneumoniae) antigens; (ii) one or more compounds of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or pharmaceutically acceptable salt(s) thereof; and (iii) a pharmaceutically acceptable carrier, wherein the one or more S. pneumoniae antigens are capable of inducing an immune response against one or more serotypes of S. pneumoniae.

[0007] The present invention also relates to formulations and compositions comprising: (i) one or more Streptococcus pneumoniae polysaccharide carrier protein conjugates, each conjugate comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the carrier protein is CRM197; (ii) one or more compounds of Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or pharmaceutically acceptable salt(s) thereof; and (iii) a pharmaceutically acceptable carrier. Furthermore, the present invention relates to the use of a formulation or composition comprising: (i) one or more Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the carrier protein is CRM197; (ii) one or more compounds of Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or pharmaceutically acceptable salt(s) thereof; and (iii) a pharmaceutically acceptable carrier, for the prevention of pneumococcal disease or for preventing or reducing the likelihood of infection with Streptococcus pneumoniae.

[0008] The present invention also relates to formulations and compositions comprising (i) one or more Streptococcus pneumoniae antigens, (ii) one or more compounds of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or pharmaceutically acceptable salts thereof, (iii) one or more sorbitan-based surfactants, and iv) one or more terpenes. In some embodiments, the one or more Streptococcus pneumoniae antigens are Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein. In some embodiments, the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.

[0009] The present invention also relates to formulations and compositions comprising: (i) one or more Streptococcus pneumoniae antigens; (ii) one or more compounds of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) sorbitan trioleate (SPAN-85); (iv) polysorbate-20 (PS-20) or polysorbate-80 (PS-80); and (v) squalene. In some embodiments, the one or more Streptococcus pneumoniae antigens are Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein. In some embodiments, the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, wherein the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.

[0010] The present invention also relates to compositions comprising the pneumococcal conjugates disclosed herein and a stable nanoemulsion "SNE." As an example, a specific SNE comprises N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piprazin-1-yl)-5-oxopentyl)stearamide, also known as "Compound A-1." This specific SNE is referred to as Compound A-1-SNE and comprises i) Compound A-1, ii) sorbitan trioleate (SPAN-85), (iii) polysorbate-20 (PS-20) or polysorbate-80 (PS-80), and (iv) squalene. As another example, a specific SNE includes N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, also known as "Compound B-1." This specific SNE is referred to as Compound B-1-SNE and includes i) Compound B-1, ii) SPAN-85, (iii) PS-20 or PS-80, and (iv) squalene. As another example, a specific SNE includes N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, also known as "Compound C-4." This particular SNE is referred to as compound C-4-SNE and includes i) compound C-4, ii) SPAN-85, (iii) PS-20 or PS-80, and (iv) squalene.

[0011] The present invention also relates to immunogenic compositions comprising a pneumococcal polysaccharide or pneumococcal conjugate and a SNE as disclosed herein.

[0012] The present invention also relates to methods for treating or preventing pneumococcal disease in a patient in need thereof by administering to the patient an immunogenic composition of the present invention.

[0013] The present invention also relates to a method for preventing or reducing the likelihood of infection by one or more serotypes of Streptococcus pneumoniae, comprising administering to a patient in need thereof an immunogenic composition of the present invention. [Brief explanation of the drawings]

[0014] [Figure 1] Example 10 shows OD640 readings after incubation of different concentrations of compound B-1-SNE with human TLR7 (top panel) or TLR8 (bottom panel) expressing HEK-Blue™ 293 cells (InvivoGen). The Y-axis represents the total compound B-1-SNE concentration. [Figure 2] Example 11 shows anti-6B IgG titers pre-immunization (day 0, pooled serum), post-dose 1 (day 21), post-dose 2 (day 42), and post-dose 3 (day 70) after immunization of mice with the formulations listed in Table 7. Error bars are geometric means with 95% confidence intervals. Transformed data analyzed by one-way ANOVA with Dunnett's post-hoc test, *p<0.05. [Figure 3] (Example 11) Pre-immune (day 0), post-dose 1 (day 21), post-dose 2 (day 42), and post-dose 3 (day 70) anti-6B OPA titers from pooled sera after immunization of mice with the formulations listed in Table 7. [Figure 4] Example 12 shows the ratios of serotype-specific IgG titers in infant rhesus macaques after immunization with PCV24 containing Compound B-1-SNE (150 μg dose of Compound B-1), PCV24 containing Compound B-1-SNE (50 μg of Compound B-1), and PCV24 containing Compound B-1-SNE (15 μg of Compound B-1) compared to immunization with PCV24 formulated without adjuvant at post-dose 3. Data for serotypes 6C and 15B are included to assess cross-reactivity. [Figure 5]Example 12 shows the ratios of serotype-specific IgG titers in infant rhesus macaques after immunization with PCV24 with Compound B-1-SNE (150 μg dose of Compound B-1), PCV24 with Compound B-1-SNE (50 μg of Compound B-1), and PCV24 with Compound B-1-SNE (15 μg of Compound B-1) compared to immunization with PCV24 formulated with APA after dose 3. Data for serotypes 6C and 15B are included to assess cross-reactivity. [Figure 6] (Example 12) Figure showing the ratio of serotype-specific IgG titers in infant rhesus macaques after immunization with PCV24 with Compound B-1-SNE (150 μg dose of Compound B-1), PCV24 with Compound B-1-SNE (50 μg of Compound B-1), and PCV24 with Compound B-1-SNE (15 μg of Compound B-1) compared to immunization with PCV20 at dose 3. [Figure 7] (Example 13) Figure showing the ratio of serotype-specific IgG titers in infant rhesus macaques after immunization with PCV24 with Compound B-1-SNE (100 μg dose of Compound B-1), PCV24 with Compound B-1-SNE (10 μg of Compound B-1), PCV24 with Compound B-1-SNE (1 μg of Compound B-1), and PCV24 with Compound B-1-SNE (0.1 μg of Compound B-1) compared to immunization with PCV20 at dose 3. [Figure 8] Example 14 shows the ratio of serotype-specific IgG titers in mice after immunization with PCV26 containing Compound B-1-SNE (10 μg dose of Compound B-1) compared to immunization with PCV26 formulated with APA at dose 2. Data for serotypes 6C and 15B are included to assess cross-reactivity. [Figure 9] Example 14 shows the IgG2a / IgG1 titer ratios of serotype-specific antibodies in mice after immunization with PCV26 containing Compound B-1-SNE (a 10 μg dose of Compound B-1) or PCV26 formulated with APA at dose 3. Responses from three representative serotypes, 18C, 19F, and 24F, are shown (left to right). [Figure 10A]Example 14: Mice immunized with PCV26 bearing Compound B-1-SNE (a 10 μg dose of Compound B-1) or PCV26 formulated with APA are protected from intratracheal challenge with Streptococcus pneumoniae 24F. Bacteremia was quantified 24 hours after infection with serotype 24F bacteria and expressed as log CFU / mL. [Figure 10B] Example 14: Mice immunized with PCV26 bearing Compound B-1-SNE (a 10 μg dose of Compound B-1) or PCV26 formulated with APA are protected from intratracheal challenge with Streptococcus pneumoniae 24F. Survival was assessed approximately 240 hours after infection with serotype 24F bacteria. DETAILED DESCRIPTION OF THE INVENTION

[0015] The present invention relates to a compound comprising one or more Streptococcus pneumoniae antigens and a compound having the structure set forth in Formula I: [ka] (In the formula, R a is selected from H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, and —NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted with 1 to 4 substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine; R a’is selected from H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, and —NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted with 1 to 4 substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine; R a’’ is selected from H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, and —NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted with 1 to 4 substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine; R' and R" are independently selected from H, (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl are optionally substituted with one to four substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; or R' and R" together with the nitrogen to which they are attached, join together to form a (C3-C6)heterocycloalkyl, wherein said (C3-C6)heterocycloalkyl is optionally substituted with one to four substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; R b is independently selected from H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted with from 1 to 4 substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine; A is (C1-C6) alkyl, (C3-C6) heterocycloalkyl, heterocycloalkyl-C(O)-R z -, (C1-C4) alkyl-N(R z )-R za carbon or nitrogen bond spacer selected from -, aryl, and heteroaryl, wherein said (C1-C6)alkyl, (C3-C6)heterocycloalkyl, aryl, and heteroaryl are independently selected from the group consisting of -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, and NR'R''; each of the (C1-C6) alkyl, (C1-C6) alkenyl, (C1-C6) alkynyl, -O(C1-C6) alkyl, -O(C1-C6) alkenyl, and -O(C1-C6) alkynyl is optionally substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine; R z each occurrence is independently H or (C1-C6) alkyl; B is [ka] is a functional group selected from D is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl, wherein the (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl is optionally substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine, or D is [ka] and Each occurrence of Z is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C20 )alkynyl, wherein the (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl is optionally substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine; m is 0, 1, 2, 3, 4, or 5; (n is 0, 1, 2, 3, 4, or 5) or a pharmaceutically acceptable salt thereof.

[0016] The present invention relates to a compound comprising one or more Streptococcus pneumoniae antigens and a compound having the structure according to formula Ia: [ka] (In the formula, R' and R" are independently selected from H, (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl are optionally substituted with one to four substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; or R' and R" together with the nitrogen to which they are attached, join together to form a (C3-C6)heterocycloalkyl, wherein said (C3-C6)heterocycloalkyl is optionally substituted with one to four substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; R beach occurrence of is -O(C1-C4)alkyl, wherein said -O(C1-C4)alkyl is optionally substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; A is, [ka] is selected from R z each occurrence is independently H or (C1-C6) alkyl; R d each occurrence of is independently selected from —OH, (C1-C4)alkyl, —O(C1-C4)alkyl, chlorine, and fluorine; B is [ka] and D is [ka] wherein any carbon on the lipid chain may be substituted with -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, or fluorine; [ka] is cis or trans stereochemistry, X 1 -O-, -C(R) 2 - or -NR-, each occurrence of R is independently selected from H, (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, —OH, —O(C-C)alkyl, —O(C-C)alkenyl, —O(C-C)alkynyl, chlorine, and fluorine; m is 0, 1, or 2; n is 0, 1, 2, or 3; p is 0, 1, or 2; q is 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; s is 1, 2, 3, 4, 5, 6, 7, or 8, t is 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18) or a pharmaceutically acceptable salt thereof.

[0017] The present invention relates to a compound comprising one or more Streptococcus pneumoniae antigens and a compound having the structure set forth in Formula II: [ka] (In the formula, R 1 is (C1-C6) alkyl, wherein said (C1-C6) alkyl is optionally substituted with 1 to 4 substituents independently selected from -OH and -O(CH3), R 2 is H, methyl or -O(CH3), R 3 each occurrence is independently H, (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, or —O(C-C)alkyl, wherein said (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, or —O(C-C)alkyl is optionally substituted with one or two substituents independently selected from —OH and —O(CH); R 4 each occurrence is independently H, (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, or —O(C-C)alkyl, wherein said (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, or —O(C-C)alkyl is optionally substituted with one or two substituents independently selected from —OH and —O(CH); R 5 teeth, [ka] and R 6 is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )alkynyl, wherein said (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl is optionally substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine; each occurrence of n is 4) or a pharmaceutically acceptable salt thereof.

[0018] The present invention relates to a compound comprising one or more Streptococcus pneumoniae antigens and a compound having the structure according to formula IIa: [ka] (In the formula, R 1 is butyl, said butyl being optionally substituted with one or two -OH groups; R 3 each occurrence of is independently H or —O(CH), R 5 teeth, [ka] and R 6 is (C 10 -C 20 ) alkyl, (C 10 -C 20 ) alkenyl, and (C 10 -C 20 )alkynyl) or a pharmaceutically acceptable salt thereof.

[0019] The present invention relates to a compound comprising one or more Streptococcus pneumoniae antigens and a compound having the structure set forth in Formula III: [ka] (In the formula, R 1 is (C1-C6) alkyl, wherein said (C1-C6) alkyl is optionally substituted with 1 to 4 substituents independently selected from -OH and -O(CH3), R 2 is H, methyl or -O(CH3), R 3 each occurrence is independently H, (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, or —O(C-C)alkyl, wherein said (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, or —O(C-C)alkyl is optionally substituted with one or two substituents independently selected from —OH and —O(CH); R 4 is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )alkynyl, wherein said (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl is optionally substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine; n is 4) or a pharmaceutically acceptable salt thereof.

[0020] The present invention relates to a compound comprising one or more Streptococcus pneumoniae antigens and a compound having the structure according to Formula IIIa: [ka] During the ceremony, (R 1 is butyl, said butyl being optionally substituted with one or two -OH groups; R 3 each occurrence of is independently H or —O(CH), R 4 is (C 10 -C 20 ) alkyl, (C 10 -C 20 ) alkenyl, and (C 10 -C 20 )alkynyl) or a pharmaceutically acceptable salt thereof.

[0021] The present invention relates to a compound comprising one or more Streptococcus pneumoniae antigens and a compound having the structure set forth in Formula IV: [ka] During the ceremony, (R 1 is (C1-C6) alkyl, wherein said (C1-C6) alkyl is optionally substituted with 1 to 4 substituents independently selected from -OH and -O(CH3), R 2 is H, methyl or -O(CH3), R 3 each occurrence is independently H, (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, or —O(C-C)alkyl, wherein said (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, and —O(C-C)alkyl are optionally substituted with one or two substituents independently selected from —OH and —O(CH); R 4 Each occurrence of (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )alkynyl, wherein the (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20) alkynyl is optionally substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, or fluorine; n is 4) or a pharmaceutically acceptable salt thereof.

[0022] The present invention relates to a compound comprising one or more Streptococcus pneumoniae antigens and a compound having the structure according to Formula IVa: [ka] (In the formula, R 1 is butyl, said butyl being optionally substituted with one or two -OH groups; R 3 each occurrence of is independently H or —O(CH), R 4 Each occurrence of (C 10 -C 20 ) alkyl, (C 10 -C 20 ) alkenyl, and (C 10 -C 20 )alkynyl) or a pharmaceutically acceptable salt thereof.

[0023] The present invention relates to a method for treating a pneumoniae infection comprising administering to a patient a therapeutically effective amount of one or more Streptococcus pneumoniae antigens in combination with one or more of the following compounds: (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, also known as compound A-1; (S)—N-(5-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, also known as compound A-2; (S)-1-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)octadecan-1-one, also known as compound A-3 N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, also known as compound B-1; N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)tetradecanamide, also known as compound B-2; N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)oleamide, also known as compound B-3; (9Z,12Z)-N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)octadeca-9,12-dienamide, also known as compound B-4; N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepan-1-yl)-5-oxopentyl)stearamide, also known as compound B-5; N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperidin-1-yl)-5-oxopentyl)stearamide, also known as compound B-6; N-(5-(3-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)azetidin-1-yl)-5-oxopentyl)stearamide, also known as compound B-7; 1-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperidine-4-carboxamide, also known as compound B-8; (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-octadecylcyclobutane-1-carboxamide, also known as compound B-9; (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-hexadecylcyclobutane-1-carboxamide, also known as compound B-10; N-(3-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-3-oxopropyl)stearamide, also known as compound B-11; N-(7-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-7-oxoheptyl)stearamide, also known as compound B-12; N-(3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)cyclobutyl)stearamide, also known as compound B-13; N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-4,4-dimethyl-5-oxopentyl)stearamide, also known as compound B-14; N-(6-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-methyl-6-oxohexan-2-yl)stearamide, also known as compound B-15; 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecyloxy)pentan-1-one, also known as compound B-16; 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecylamino)pentan-1-one, also known as compound B-17; N-(5-(4-(4-((5-amino-7-(butylamino)-3-methyl-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, also known as compound B-18; (9Z,12Z)-N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)octadeca-9,12-dienamide, also known as compound C-1; N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)tetradecanamide, also known as compound C-2 N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)oleamide, also known as compound C-3 N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, also known as compound C-4; N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)-4-oxobutyl)stearamide, also known as compound C-5; (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)carbamate, also known as compound D-1; 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperazine-1-carboxamide, also known as compound D-2; 3-stearamidopropyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazine-1-carboxylate, also known as compound D-3 N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-6,6,6-trifluorohexyl)stearamide, also known as compound D-4; N-(4-((4-((7-(butylamino)-5-hydroxy-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, also known as compound D-5, and (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)carbamate, also known as compound D-6; or a pharmaceutically acceptable salt thereof.

[0024] The present invention also provides a composition comprising one or more Streptococcus pneumoniae antigens and a stable nanoemulsion, wherein the stable nanoemulsion comprises (i) one or more compounds having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt(s) thereof, ii) one or more emulsifying agents, and iii) one or more terpenes.

[0025] The present invention also provides a composition comprising one or more Streptococcus pneumoniae antigens and a stable nanoemulsion, wherein the stable nanoemulsion comprises (i) one or more compounds having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt(s) thereof, ii) one or more sorbitan-based surfactants, and iii) one or more terpenes.

[0026] The present invention also provides a composition comprising one or more Streptococcus pneumoniae antigens and a stable nanoemulsion, wherein the stable nanoemulsion comprises (i) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof, (ii) SPAN-85, (iii) PS-20 or PS-80, and (iv) squalene.

[0027] The present invention also provides a composition comprising one or more Streptococcus pneumoniae antigens and a stable nanoemulsion, wherein the stable nanoemulsion comprises (i) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof, (ii) SPAN-85, (iii) PS-20 or PS-80, and (iv) squalene.

[0028] The present invention also provides pharmaceutical compositions comprising: (i) at least one Streptococcus pneumoniae (S. pneumoniae) polysaccharide; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0029] The present invention also provides a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae (S. pneumoniae) polysaccharide; (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0030] The present invention also provides an immunogenic composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0031] The present invention also provides an immunogenic composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide; (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0032] The present invention also provides a single-dose vaccine composition comprising: (i) at least one pneumococcal polysaccharide; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; and (iv) PS-20 or PS-80, wherein a single dose of the vaccine composition is sufficient to elicit a desired immune response against the at least one pneumococcal polysaccharide.

[0033] The present invention also provides a single-dose vaccine composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide; (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier, wherein a single dose of the vaccine composition is sufficient to elicit a desired immune response against the at least one Streptococcus pneumoniae polysaccharide.

[0034] The present invention also provides a method for treating or preventing pneumococcal disease in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0035] The present invention also provides a method for treating or preventing pneumococcal disease in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide; (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0036] The present invention also provides pharmaceutical compositions comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0037] The present invention also provides pharmaceutical compositions comprising (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier.

[0038] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharides being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A , 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 1 9A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22 The present invention provides a pharmaceutical composition comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a serotype selected from the group consisting of F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM197; (ii) a compound having a structure according to Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0039] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharides being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; , 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 2 2F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. The present invention provides a pharmaceutical composition comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a serotype selected from the group consisting of F, 24F, 33F, and 35B, wherein the carrier protein is CRM197; (ii) a compound having a structure according to Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0040] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharides being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; A) De-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. 3F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) from the group of serotypes selected from 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. wherein the carrier protein is CRM197; (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0041] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharides are selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; -acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35 B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is C. Provided is a pharmaceutical composition comprising a Streptococcus pneumoniae polysaccharide carrier protein conjugate that is RM197; (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0042] The present invention also provides immunogenic compositions comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0043] The present invention also provides immunogenic compositions comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0044] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharides being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A , 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19 A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, The present invention provides an immunogenic composition comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a serotype selected from the group consisting of 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM197; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0045] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharide being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F , 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. The present invention provides an immunogenic composition comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a serotype selected from the group consisting of 4F, 33F, and 35B, wherein the carrier protein is CRM197; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0046] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharides being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; A) De-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33 F, and 35B; e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. and (v) squalene.

[0047] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharides are selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; -acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B , e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM. (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0048] The present invention also provides a single-dose vaccine composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene, wherein a single dose of the vaccine composition is sufficient to elicit a desired immune response against the at least one polysaccharide carrier protein conjugate.

[0049] The present invention also provides a single-dose vaccine composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0050] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharides being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; , de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, and 33F; and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, (ii) a compound having a structure according to Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene, wherein a single dose of the vaccine composition is sufficient to elicit a desired immune response against the polysaccharide carrier protein conjugate.

[0051] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharides are selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; Acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM197. (ii) a compound having a structure according to Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier, wherein a single dose of the vaccine composition is sufficient to elicit a desired immune response against the polysaccharide carrier protein conjugate.

[0052] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of which comprises a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharides being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and e) 1, 3, 4, 5, 6A, 6B, 7F and f) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a serotype selected from the group consisting of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM197. (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene, wherein a single dose of the vaccine composition is sufficient to elicit a desired immune response against the polysaccharide carrier protein conjugate.

[0053] The present invention also provides (i) Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharides being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; 9A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 1 (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of the group of serotypes selected from 1A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of the group of serotypes selected from 1A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM197; and Provided is a single-dose vaccine composition comprising 4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier, wherein a single dose of the vaccine composition is sufficient to elicit a desired immune response against the polysaccharide carrier protein conjugate.

[0054] The present invention also provides a method for treating or preventing pneumococcal disease in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0055] The present invention also provides a method for treating or preventing pneumococcal disease in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0056] The present invention also provides a method of treating or preventing pneumococcal disease in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharide being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16 F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A , 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM197; (ii) a compound having a structure according to Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0057] The present invention also relates to a method of treating or preventing pneumococcal disease in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharide being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; , 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19 A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F , 33F, and 35B, wherein the carrier protein is CRM197; (ii) a compound having a structure according to Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0058] The present invention also relates to a method of treating or preventing pneumococcal disease in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharide being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 2 a) a group of serotypes selected from the group consisting of serotypes selected from the group consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; b) a group of serotypes selected from the group consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; The method includes administering to a patient a pharmaceutical composition comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, wherein the carrier protein is CRM197; (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0059] The present invention also relates to a method of treating or preventing pneumococcal disease in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharide being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33 F, and 35B; e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM197. (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0060] The present invention also provides a method for preventing or reducing the likelihood of infection with pneumococcus in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0061] The present invention also provides a method for preventing or reducing the likelihood of infection with pneumococcus in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having a structure set forth in Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0062] The present invention also relates to a method for preventing or reducing the likelihood of infection with Streptococcus pneumoniae in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharide being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 2 The method includes administering to a patient a pharmaceutical composition comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a serotype selected from the group consisting of 3A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM197; (ii) a compound having a structure according to Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0063] The present invention also relates to a method for preventing or reducing the likelihood of infection with Streptococcus pneumoniae in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharide being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; , 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C , 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24 The method includes administering to a patient a pharmaceutical composition comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a serotype selected from the group consisting of F, 33F, and 35B, wherein the carrier protein is CRM197; (ii) a compound having a structure according to Formula I, Ia, II, IIa, III, IIIa, IV or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0064] The present invention also relates to a method for preventing or reducing the likelihood of infection with Streptococcus pneumoniae in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharide being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23 B, 23F, 24F, 33F, and 35B; e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, wherein the carrier protein is CRM197; (iii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iv) SPAN-85; (v) PS-20 or PS-80; and (vi) squalene.

[0065] The present invention also relates to a method for preventing or reducing the likelihood of infection with Streptococcus pneumoniae in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae polysaccharide being selected from the group consisting of: a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; , 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24 F, 33F, and 35B; e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM1. 97; (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0066] In specific embodiments of the above i) pharmaceutical compositions, ii) immunogenic compositions, and iii) single-dose vaccine compositions, the compositions comprise more than twenty (20) different Streptococcus pneumoniae polysaccharide carrier protein conjugates. For example, the present invention provides (i) a pharmaceutical composition comprising a Streptococcus pneumoniae polysaccharide carrier protein conjugate comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide carrier protein conjugate comprises a Streptococcus pneumoniae polysaccharide from a group of serotypes selected from more than twenty (20) different Streptococcus pneumoniae polysaccharides.

[0067] In the above specific embodiments, the group of serotypes is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; or b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A , de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

[0068] In the above specific embodiments, the group of serotypes is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; or b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated or c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.

[0069] In specific embodiments of i) the pharmaceutical composition, ii) the immunogenic composition, and iii) the single-dose vaccine composition above, the composition comprises more than twenty-five (25) different Streptococcus pneumoniae polysaccharide carrier protein conjugates. For example, the present invention provides (i) a pharmaceutical composition comprising a Streptococcus pneumoniae polysaccharide carrier protein conjugate comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide carrier protein conjugate comprises a Streptococcus pneumoniae polysaccharide from a group of serotypes selected from more than twenty-five (25) different Streptococcus pneumoniae polysaccharides.

[0070] In the above specific embodiments, the group of serotypes is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; or b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A , de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

[0071] In the above specific embodiments, the group of serotypes is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B; or b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated or c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.

[0072] In some embodiments of the above i) pharmaceutical compositions, ii) immunogenic compositions, and iii) single-dose vaccine compositions, the concentration of the compound is 0.01 μg / mL to 1000 μg / mL, or 0.1 μg / mL to 100 μg / mL, or 80 μg / mL, or 16 μg / mL, or 4 μg / mL.

[0073] In some embodiments of the above i) pharmaceutical compositions, ii) immunogenic compositions, and iii) single-dose vaccine compositions, the concentration of SPAN-85 is 0.001 mg / mL to 100 mg / mL, or 0.01 mg / mL to 50 mg / mL, or 0.1 mg / mL to 10 mg / mL.

[0074] In some embodiments of the above i) pharmaceutical compositions, ii) immunogenic compositions, and iii) single-dose vaccine compositions, the concentration of PS-20 or PS-80 is 0.001 mg / mL to 100 mg / mL, or 0.01 mg / mL to 50 mg / mL, or 0.1 mg / mL to 10 mg / mL.

[0075] In some embodiments of the above i) pharmaceutical compositions, ii) immunogenic compositions, and iii) single-dose vaccine compositions, the concentration of squalene is 0.01 mg / mL to 100 mg / mL, or 0.02 mg / mL to 20 mg / mL, or 1 mg / mL to 20 mg / mL.

[0076] In some embodiments of the above i) pharmaceutical composition, ii) immunogenic composition, and ii) single-dose vaccine composition, the concentration of the compound is 80 μg / mL, the concentration of SPAN-85 is 4.8 mg / mL, the concentration of PS-20 or PS-80 is 4.8 mg / mL, and the concentration of squalene is 16 mg / mL.

[0077] In some embodiments of i) the pharmaceutical composition, ii) the immunogenic composition, and ii) the single-dose vaccine composition described above, and iv) the single-dose vaccine composition described above, the concentration of the compound is 16 μg / mL, the concentration of SPAN-85 is 4.8 mg / mL, the concentration of PS-20 or PS-80 is 4.8 mg / mL, and the concentration of squalene is 16 mg / mL.

[0078] In some embodiments of i) the pharmaceutical composition, ii) the immunogenic composition, and ii) the single-dose vaccine composition described above, and iv) the single-dose vaccine composition described above, the concentration of the compound is 4 μg / mL, the concentration of SPAN-85 is 4.8 mg / mL, the concentration of PS-20 or PS-80 is 4.8 mg / mL, and the concentration of squalene is 16 mg / mL.

[0079] In some embodiments of the above i) pharmaceutical compositions, ii) immunogenic compositions, and iii) single-dose vaccine compositions, the compositions further comprise L-met and EDTA.

[0080] Abbreviations and Definitions The following abbreviations are used herein:

[0081] [Table 1] TIFF2026505682000018.tif230151TIFF2026505682000019.tif227136TIFF2026505682000020.tif204152

[0082] As used throughout this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0083] As used herein, the term "about," when used herein in reference to a value, refers to a value that is the same as the referenced value or, in context, is similar to the referenced value. Generally, a person skilled in the art, familiar with the context, will understand the absolute amount and / or relative degree of difference encompassed by "about" in that context. For example, in some embodiments, the term "about" can encompass a range of values ​​within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less of the referenced value.

[0084] As defined herein, "adjuvant" refers to a compound or compound formulation or composition that serves to enhance the immunogenicity of the compositions of the invention. An adjuvant may i) enhance the immune response to an antigen (e.g., pneumococcal polysaccharide) that is weakly immunogenic when administered alone, e.g., induces no or a weak antibody titer or cell-mediated immune response, ii) increase the antibody titer against the antigen, and / or iii) reduce the dose of the antigen effective to achieve an immune response in an individual.

[0085] As used herein, the term "administration" refers to the act of providing an active agent, composition, or formulation to a subject. Exemplary routes of administration to the human body can be by eye (ophthalmic), mouth (oral), skin (transdermal), nose (intranasal), lung (inhalant), rectum, vagina, oral mucosa (buccal), ear, injection (e.g., intravenous (IV), subcutaneous, intratumoral, intraperitoneal, intramuscular (IM), intradermal (ID), etc.), etc.

[0086] As used herein, "agent" refers to any chemical class of particle, compound, molecule, or entity.

[0087] As used herein, the term "alkyl" refers to a straight-chain, cyclic, or branched saturated aliphatic hydrocarbon having a specified number of carbon atoms. Numerical ranges may be given, referring to the total chain length. For example, a C1-C6 alkyl has a chain length of 1 to 6 atoms. Alkyl groups may be substituted with one or more "substituents," which may be the same or different and are as defined herein below. Unless otherwise indicated, alkyl groups are unsubstituted.

[0088] As used herein, the term "alkenyl" refers to a straight-chain, cyclic, or branched unsaturated aliphatic hydrocarbon having the specified number of carbon atoms, including, but not limited to, diene, triene, and tetraene unsaturated aliphatic hydrocarbons. Alkenyl groups can be substituted with one or more "substituents," which may be the same or different, and are as defined herein below. Unless otherwise specified, an alkenyl group is unsubstituted.

[0089] As used herein, the term "alkynyl" refers to a straight-chain, cyclic, or branched unsaturated aliphatic hydrocarbon having the specified number of carbon atoms, including, but not limited to, diene, triene, and tetraene unsaturated aliphatic hydrocarbons. Alkynyl groups can be substituted with one or more "substituents," which may be the same or different, and are as defined herein below. Unless otherwise indicated, an alkynyl group is unsubstituted.

[0090] As used herein, the term "antigen" refers to any antigen capable of generating one or more immune responses. An antigen can be a protein, peptide, or polypeptide. In certain embodiments, an antigen is a lipid or carbohydrate. In certain embodiments, an antigen is a polysaccharide. In certain embodiments, an antigen is a pneumococcal polysaccharide. In certain embodiments, a polysaccharide is a Streptococcus pneumoniae polysaccharide. An antigen can be one that generates a humoral and / or CTL immune response.

[0091] As used herein, the term "aryl" refers to a carbocyclic aromatic monocyclic or bicyclic ring system containing from about 6 to about 14 carbon atoms. In one embodiment, an aryl group contains from about 6 to about 10 carbon atoms. Aryl groups can be substituted with one or more "ring system substituents," which may be the same or different, and are as defined herein below. Non-limiting examples of aryl groups include phenyl and naphthyl. In one embodiment, an aryl group is phenyl. Unless otherwise indicated, an aryl group is unsubstituted.

[0092] As used herein, the term "composition" refers to a formulation containing an active pharmaceutical ingredient or biological component (e.g., a pneumococcal polysaccharide carrier protein conjugate and a compound) along with one or more additional ingredients. The term "composition" is used interchangeably with "pharmaceutical composition" and "formulation." The composition may be liquid or solid (e.g., lyophilized). Additional components that may be optionally included include pharmaceutically acceptable excipients, additives, diluents, buffers, sugars, amino acids, chelating agents, surfactants, polyols, bulking agents, stabilizers, cryoprotectants, solubilizers, emulsifiers, salts, adjuvants, tonicity agents, delivery vehicles, and antimicrobial preservatives. The composition is nontoxic to recipients at the dosages and concentrations employed.

[0093] As used herein, the term "comprising" when used in connection with the compositions of the present invention refers to the inclusion of any other ingredients such as adjuvants and excipients, or the addition of one or more polysaccharide carrier protein conjugates not specifically listed.

[0094] As used herein, the term "consisting of" and variations such as "consist of" when used in conjunction with a polyvalent polysaccharide mixture or a polyvalent polysaccharide carrier protein conjugate mixture refers to a mixture having those particular pneumococcal polysaccharides or pneumococcal polysaccharide carrier protein conjugates and not having other pneumococcal polysaccharides or pneumococcal polysaccharide carrier protein conjugates from different serotypes.

[0095] As used herein, the term "consists essentially of" and variations such as "consist essentially of" or "consisting essentially of" refer to the inclusion of any recited element or group of elements, and the optional inclusion of other elements of similar or different nature to the recited elements, which do not materially alter the basic or novel characteristics of the specified dosing regimen, method, or composition.

[0096] As used herein, the term "de-O-acetylated-15B" or "de-O-acetyl-15B" or "de-O-Ac-15B" refers to de-O-acetylated serotype 15B having an O-acetyl content of less than 10% per repeat unit. In another embodiment, the O-acetyl content is less than 5% per repeat unit. In another embodiment, the O-acetyl content is less than about 1% per repeat unit. In another embodiment, the O-acetyl content is less than 1% per repeat unit. In another embodiment, the O-acetyl content is less than 0.5% per repeat unit. In another embodiment, the O-acetyl content is less than 0.1% per repeat unit. In another embodiment, the O-acetyl content is 0% per repeat unit. The process of de-O-acetylation is known in the art, for example, as described in Rajam et al., Clinical and Vaccine Immunology, 2007, 14(9):1223-1227.

[0097] As used herein, the term "dose" means the amount of a drug, API (active pharmaceutical ingredient), formulation, composition, pharmaceutical composition, or immunogenic composition that is taken or recommended to be taken at a particular time.

[0098] As used herein, the term "heteroaryl" refers to an aromatic monocyclic or polycyclic ring system containing about 5 to about 14 ring atoms, wherein 1 to 4 of the ring atoms are independently O, N, or S, and the remaining ring atoms are carbon atoms. In one embodiment, a heteroaryl group has 5 to 10 ring atoms. In another embodiment, a heteroaryl group is monocyclic and has 5 or 6 ring atoms. In another embodiment, a heteroaryl group is bicyclic. A heteroaryl group may be substituted with one or more "ring system substituents," which may be the same or different, and are as defined herein below. A heteroaryl group is joined via a ring carbon atom, and any nitrogen atom of a heteroaryl may be oxidized to the corresponding N-oxide. In one embodiment, a heteroaryl group is a 5-membered heteroaryl. In another embodiment, a heteroaryl group is a 6-membered heteroaryl. In another embodiment, a heteroaryl group includes a 5- to 6-membered heteroaryl group fused to a benzene ring. Unless otherwise indicated, a heteroaryl group is unsubstituted.

[0099] As used herein, the term "heterocycloalkyl" refers to a saturated or partially unsaturated non-aromatic monocyclic, bicyclic (including spirocyclic), or bridged carbocyclic ring or ring system containing 3 to about 11 ring atoms, at least one ring heteroatom selected from N, S, and O, with the remaining ring atoms being carbon atoms. A heterocycloalkyl group can be attached through a ring carbon or a ring nitrogen atom, unless otherwise specified. A heterocycloalkyl ring can be substituted on a ring carbon and / or one or more ring nitrogens. In one embodiment, a heterocycloalkyl group is monocyclic and has about 3 to about 7 ring atoms. In another embodiment, a heterocycloalkyl group is monocyclic and has about 4 to about 7 ring atoms. In other embodiments, a heterocycloalkyl group is bicyclic and has 7 to 10 ring atoms, 8 to 10 ring atoms, or 9 or 10 ring atoms. In yet another embodiment, a heterocycloalkyl group is monocyclic and has 5 or 6 ring atoms. In one embodiment, a heterocycloalkyl group is monocyclic. In another embodiment, the heterocycloalkyl group is bicyclic. The heterocycloalkyl group can be substituted. In some embodiments, the heterocycloalkyl group has one to two heteroatoms in the ring selected from nitrogen, sulfur, and oxygen atoms. In some embodiments, the heterocycloalkyl group has one heteroatom in the ring selected from nitrogen, sulfur, and oxygen atoms. In some embodiments, the heteroatom is selected from O, S, S(O), S(O)2, and -NH-, -N(alkyl)-. Non-limiting examples include aliphatic groups containing heteroatoms, such as ethers, thioethers, amines, hydroxymethyl, 3-hydroxypropyl, 1,2-dihydroxyethyl, 2-methoxyethyl, 2-aminoethyl, and 2-dimethylaminoethyl.

[0100] As used herein, the terms "immunogenic" or "immunogenicity" refer to the ability of an antigen (e.g., Streptococcus pneumoniae polysaccharide) to elicit an immune response in a subject. The term "immunogenic composition" refers to the ability of a drug, API, formulation, composition, or pharmaceutical composition to elicit an immune response in a subject.

[0101] As used herein, the phrase patient or subject "in need of treatment" includes those who have been previously exposed to or infected with S. pneumoniae polysaccharide, those who have previously been vaccinated against S. pneumoniae polysaccharide, as well as anyone who is susceptible to infection or in whom a reduction in the likelihood of infection is desired, such as an immunocompromised, elderly, child, adult, or healthy individual.

[0102] As used herein, the phrase "indicated for the prevention of pneumococcal disease" means that the vaccine or composition has been approved by one or more regulatory authorities, such as the U.S. Food and Drug Administration, for the prevention of one or more diseases caused by any serotype of Streptococcus pneumoniae, including, but not limited to, pneumococcal disease generally, invasive pneumococcal disease (IPD), pneumococcal pneumonia (PP), pneumococcal meningitis, pneumococcal bacteremia, invasive disease caused by Streptococcus pneumoniae, and otitis media caused by Streptococcus pneumoniae.

[0103] As used herein, the term "multiple dose" refers to a vaccine composition, or pharmaceutical composition, or immunogenic composition that requires administration or injection of more than one dose or components therein in a clinical regimen to induce a sustained immune response, provide protection from disease, or reduce the likelihood of infection by an infectious agent. Those skilled in the art will understand how to determine a durable immune response, for example, by measuring antibody titers over a specified period of time.

[0104] As used herein, a "patient" (alternatively referred to herein as a "subject") refers to a mammal susceptible to infection with Streptococcus pneumoniae polysaccharides. In a preferred embodiment, the patient is a human. The patient can be treated prophylactically or therapeutically. Prophylactic treatment provides sufficient protective immunity to reduce the likelihood or severity of pneumococcal infection or its effects, e.g., pneumococcal pneumonia. Therapeutic treatment can be carried out to reduce the severity of Streptococcus pneumoniae infection or prevent recurrence or its clinical effects. Prophylactic treatment can be carried out using the pneumococcal conjugate compositions or vaccines, or immunogenic compositions of the invention described herein. The pneumococcal conjugate compositions or vaccines, or immunogenic compositions of the invention can be administered to the general population or to individuals at high risk of pneumococcal infection, such as the elderly, or individuals living with or caring for the elderly.

[0105] As used herein, the term "PCV1" refers to a monovalent pneumococcal conjugate vaccine or composition comprising one pneumococcal polysaccharide carrier protein conjugate comprising a capsular polysaccharide from a Streptococcus pneumoniae serotype conjugated to a carrier protein. In a specific embodiment, the carrier protein is CRM197.

[0106] As used herein, the term "PCV24" refers to a 24-valent pneumococcal conjugate vaccine or composition comprising 23 pneumococcal polysaccharide carrier protein conjugates, each comprising a capsular polysaccharide from a Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae serotypes are 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 18C, 19A, 19F, 22F, 23B, 24F, 31, 33F, and 35B, and at least one of the following serotype 15 serotypes: 15B, 15C, or de-O-acetylated-15B. In a specific embodiment, the serotype 15 serotype is serotype 15C or de-O-acetylated-15B. In another embodiment, the serotype 15 serotype is serotype de-O-acetylated-15B. In a specific embodiment, the carrier protein of one or more of the Streptococcus pneumoniae polysaccharide carrier protein conjugates is CRM 197. In a further embodiment, the carrier protein of each of the Streptococcus pneumoniae polysaccharide carrier protein conjugates is CRM 197.

[0107] As used herein, the term "PCV26" refers to a 26-valent pneumococcal conjugate vaccine or composition comprising 25 pneumococcal polysaccharide carrier protein conjugates, each comprising a capsular polysaccharide from a Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae serotypes are 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 24F, 31, 33F, and 35B, and at least one of the following serotype 15 serotypes: 15B, 15C, or de-O-acetylated-15B. In a specific embodiment, the serotype 15 serotype is serotype 15C or de-O-acetylated 15B. In another embodiment, the serotype 15 serotype is serotype de-O-acetylated 15B. In a specific embodiment, one or more carrier proteins of the Streptococcus pneumoniae polysaccharide carrier protein conjugates is CRM197. In a further embodiment, each carrier protein of the Streptococcus pneumoniae polysaccharide carrier protein conjugate is CRM197.

[0108] As used herein, the term "pharmaceutically acceptable" with respect to a carrier, diluent, or excipient in a pharmaceutical composition indicates that the carrier, diluent, or excipient must be compatible with the other ingredients of the composition and not deleterious to the recipient thereof.

[0109] As used herein, the term "pharmaceutical composition" refers to a composition containing an active pharmaceutical or biological component together with one or more additional ingredients, e.g., a composition in which an active agent is formulated with one or more pharmaceutically acceptable carriers. As used herein, the terms "pharmaceutical formulation" and "formulation" are used interchangeably with "pharmaceutical composition." In some embodiments, the active agent is present in a unit dosage amount suitable for administration in a therapeutic regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. A pharmaceutical composition or formulation may be liquid or solid (e.g., lyophilized). Additional components that may be optionally included include pharmaceutically acceptable excipients, additives, diluents, buffers, sugars, amino acids, chelating agents, surfactants, polyols, bulking agents, stabilizers, cryoprotectants, solubilizers, emulsifiers, salts, adjuvants, tonicity agents, delivery vehicles, and antimicrobial preservatives. The pharmaceutical composition or formulation is nontoxic to recipients at the dosages and concentrations used. In some embodiments, the pharmaceutical composition can be specially formulated for administration in solid or liquid form, including those suitable for: oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., buccal, sublingual, and those targeted for systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., sterile solutions or suspensions, or sustained-release formulations; topical application, e.g., as a cream, ointment, or controlled-release patch or spray applied to the skin, lungs, or oral cavity; vaginal or rectal administration, e.g., as a pessary, cream, or foam; sublingually; ophthalmically; transdermally; or intranasally, via the lungs, and other mucosal surfaces. In some embodiments, the term formulation refers to a single dose of vaccine, which can be contained in any volume suitable for injection.

[0110] As used herein, the term "pneumococcal conjugate" or "pneumococcal polysaccharide carrier protein conjugate" refers to a Streptococcus pneumoniae polysaccharide carrier protein conjugate.

[0111] As used herein, the term "pneumococcal conjugate vaccine" (or "PCV") is a pharmaceutical formulation or composition comprising one or more pneumococcal polysaccharide carrier protein conjugates that provide active immunity to a disease or pathological condition caused by one or more serotypes of Streptococcus pneumoniae.

[0112] As used herein, the term "ring system substituent" refers to a substituent attached to an aromatic or non-aromatic ring system that, for example, replaces an available hydrogen on the ring system. Ring system substituents can be the same or different and are each independently selected. Examples of ring system substituents include alkyl, alkenyl, alkynyl, aryl, heteroaryl, -OH, hydroxyalkyl, haloalkyl, -O-alkyl, -O-haloalkyl, -alkylene-O-alkyl, -O-aryl, -O-alkylene-aryl, acyl, -C(O)-aryl, halo, -NO, -CN, -SF, -C(O)OH, -C(O)O-alkyl, -C(O)O-aryl, -C(O)O-alkylene-aryl, -S(O)-alkyl, -S(O)-aryl. Examples of ring system substituents include alkyl, -S(O)-aryl, -S(O)-aryl, -S(O)-heteroaryl, -S(O)-heteroaryl, -S-alkyl, -S-aryl, -S-heteroaryl, -S-alkylene-aryl, -S-alkylene-heteroaryl, -S(O)-alkylene-aryl, -S(O)-alkylene-heteroaryl, cycloalkyl, heterocycloalkyl, -OC(O)-alkyl, -OC(O)-aryl, and -OC(O)-cycloalkyl. Further examples of ring system substituents include (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, -OH, -O(C-C)alkyl, -O(C-C)alkenyl, -O(C-C)alkynyl, chlorine, and fluorine.

[0113] As used herein, the term "single dose" refers to a vaccine composition that requires only one administration or injection in a clinical regimen to induce a durable immune response and provide protection from disease (e.g., pneumococcal disease). One of skill in the art will understand how to determine a durable immune response, for example, by measuring antibody titers over a specified period of time.

[0114] As used herein, the terms "substituent" or "optional substituent" or "optionally substituted" refer to, for example, a substituent attached to an alkyl group, or an alkenyl group, or an alkynyl group that replaces an available hydrogen on the group. The substituents can be the same or different and are each independently selected. Examples of substituents include alkyl, alkenyl, alkynyl, aryl, heteroaryl, -OH, hydroxyalkyl, haloalkyl, -O-alkyl, -O-haloalkyl, -alkylene-O-alkyl, -O-aryl, -O-alkylene-aryl, acyl, -C(O)-aryl, halo, -NO, -CN, -SF, -C(O)OH, -C(O)O-alkyl, -C(O)O-aryl, -C(O)O-alkylene-aryl, -S(O)-alkyl, -S(O)-alkyl.

[0023] Examples of substituents include -C(O)-aryl, -S(O)-aryl, -S(O)-heteroaryl, -S(O)-heteroaryl, -S-alkyl, -S-aryl, -S-heteroaryl, -S-alkylene-aryl, -S-alkylene-heteroaryl, -S(O)-alkylene-aryl, -S(O)-alkylene-heteroaryl, cycloalkyl, heterocycloalkyl, -OC(O)-alkyl, -OC(O)-aryl, and -OC(O)-cycloalkyl. Further examples of substituents include (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, -OH, -O(C-C)alkyl, -O(C-C)alkenyl, -O(C-C)alkynyl, chlorine, and fluorine.

[0115] As used herein, "SNE" or "stable nanoemulsion" refers to a composition comprising a compound of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof. In some embodiments, the SNE comprises a compound of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, sorbitan trioleate (SPAN-85), polysorbate-20 (PS-20) or polysorbate-80 (PS-80), and squalene, wherein the composition is in the form of a nanoemulsion. In some embodiments, the SNE comprises a compound of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, SPAN-85, PS-20, and squalene. In some embodiments, the SNE comprises 0.001 mg / mL to 10 mg / mL of SPAN-85, 0.001 mg / mL to 10 mg / mL of PS-20, and 0.01 mg / mL to 100 mg / mL of squalene. In further embodiments, the SNE comprises 0.05 mg / mL to 5 mg / mL of SPAN-85, 0.05 mg / mL to 5 mg / mL of PS-20, and 0.05 mg / mL to 50 mg / mL of squalene. In yet a further embodiment, the SNE comprises 0.1 mg / mL to 1 mg / mL of SPAN-85, 0.1 mg / mL to 1 mg / mL of PS-20, and 1 mg / mL to 10 mg / mL of squalene. As used herein, SNE is used interchangeably with "squalene emulsion."

[0116] Nanoemulsions, also known as nanometer-sized emulsions, are fine oil-in-water (o / w) dispersions of two immiscible fluids. Nanoemulsions are colloidal particle systems in the submicron size range that act as carriers for drug molecules. Their size ranges from 10 to 1,000 nm. These carriers are solid spheres with amorphous, lipophilic surfaces.

[0117] As used herein, the term "therapeutically effective amount" refers to an amount of an active component (antigen) sufficient to produce a desired therapeutic effect in a human or animal, e.g., the amount necessary to induce an immune response, treat, cure, prevent the onset and progression of, or inhibit a disease or its symptoms, and / or the amount necessary to ameliorate symptoms or cause regression of the disease. The therapeutically effective amount may vary depending on the structure and potency of the active component and the intended mode of administration. One skilled in the art can easily determine the therapeutically effective amount of a given active component in a vaccine.

[0118] As used herein, the term "valency" refers to the presence of a specified number of polysaccharides or polysaccharide carrier protein conjugates in a composition.

[0119] As used herein, the term "vaccine" or "vaccine composition" refers to a biological preparation used to stimulate the production of antibodies and provide immunity against infectious diseases.

[0120] As used herein, the term "carbon or nitrogen linked spacer" refers to any chemistry that links a carbon (-C) or nitrogen (-N) to a benzyl group in the formulae of the present invention (see the bond in variable "A" in Formulae I and Ia). Examples of carbon or nitrogen linked spacers are C1-C6 alkyl, heterocycloalkyl, aryl, and heteroaryl, wherein said alkyl, heterocycloalkyl, aryl, and heteroaryl may be substituted.

[0121] Unless otherwise specified, when describing linkers throughout this disclosure, the first available bond on the linker group connects to the left the portion of the compound adjacent to the linker group, and the terminal available bond of the linker group connects to the right the terminal portion of the compound adjacent to the linker group. For example, if the linker (L) is -CH2-CF2-, the definition of L includes only A-CH2-CF2-B, but not A-CF2-CH2-B.

[0122] As used herein, the term "functional group" refers to any chemical entity that connects a carbon or nitrogen linking spacer to a lipid group. Examples of functional groups include: [ka] where n is 0, 1, 2, 3, 4, or 5.

[0123] Further examples of functional groups include: [ka] is.

[0124] As used herein, the term "lipid" refers to any chemical entity that is insoluble in water but soluble in organic solvents. Examples of lipids are represented by the variable D (see Formulas I and Ia) and include (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl, wherein said alkyl, alkenyl, and alkynyl may be substituted.

[0125] Further examples of lipids include: [ka] and Any carbon on the lipid chain may be substituted, [ka] is cis or trans stereochemistry, X is O, C(R) 2 , or NR, R is independently selected from H, (C1-C4) alkyl, (C1-C4) alkenyl, (C1-C4) alkynyl, OH, O(C1-C4) alkyl, O(C1-C4) alkenyl, O(C1-C4) alkynyl, chlorine, and fluorine.

[0126] Further examples of lipids are represented by the variable D (see Formulas I and Ia), where D is [ka] wherein Z is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )alkynyl, wherein said (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) The alkynyl may be substituted.

[0127] Compounds of the present invention having one or more asymmetric centers may exist as mixtures of stereoisomers or as substantially pure individual diastereomers or enantiomers, unless otherwise specified. The present invention encompasses all stereoisomeric forms of the compounds of Formulas I, Ia, II, IIa, III, IIIa, IV, and IVa. Unless a specific stereochemistry is indicated, the present invention is meant to include all such isomers of these compounds. Any asymmetric centers present in the compounds of Formulas I, Ia, II, IIa, III, IIIa, IV, and IVa can have either the (R) or the (S) configuration, independently of one another. When a bond to a chiral carbon is shown as a straight line in a structural formula of the present invention, it is understood that both the (R) and (S) configurations of the chiral carbon, and thus both enantiomers and mixtures thereof, are encompassed by the formula. Similarly, when a compound name is listed without a chiral designation for the chiral carbon, it is understood that both the (R) and (S) configurations of the chiral carbon, and thus individual enantiomers, diastereomers, and mixtures thereof, are encompassed by the name. The production of specific stereoisomers or mixtures thereof may be identified in the examples where such stereoisomers or mixtures were obtained, but this in no way limits the scope of the invention to the inclusion of all stereoisomers and mixtures thereof.

[0128] When any variable (e.g., n, Rb, etc.) occurs more than one time in any constituent or in formula I, Ia, II, IIa, III, IIIa, IV, and IVa, its definition on each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0129] A "stable" compound is one that can be prepared and isolated and whose structure and properties remain or remain essentially unchanged for a period of time sufficient to permit use of the compound for the purposes described herein (e.g., use as an adjuvant in an immunogenic composition administered to a subject). The compounds of the present invention are limited to stable compounds encompassed by Formula I, Ia II, IIa, III, IIIa, IV, and IVa.

[0130] Wavy lines as used herein [ka] indicates the point of attachment to the rest of the compound. Lines drawn through ring systems, e.g.: [ka] indicates that the bond may be attached to any of the substitutable ring atoms.

[0131] compound The compounds described herein are useful in adjuvant formulations of the invention that are useful for boosting the immunological response of a pneumococcal composition or pneumococcal conjugate composition or pneumococcal vaccine (PV) or pneumococcal conjugate vaccine (PCV) described herein.

[0132] In some embodiments, a composition of the invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I: [ka] (In the formula, R a is selected from H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, and —NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted with 1 to 4 substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine; R a’ is selected from H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, and —NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted with 1 to 4 substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine; R a’’ is selected from H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, and —NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted with 1 to 4 substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine; R' and R" are independently selected from H, (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl are optionally substituted with one to four substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; or R' and R" together with the nitrogen to which they are attached, join together to form a (C3-C6)heterocycloalkyl, wherein said (C3-C6)heterocycloalkyl is optionally substituted with one to four substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; R b is independently selected from H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted with from 1 to 4 substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine; A is (C1-C6) alkyl, heterocycloalkyl, heterocycloalkyl-C(O)-R z -, (C1-C4) alkyl-N(R z )-R za carbon or nitrogen bond spacer selected from -, aryl, and heteroaryl, wherein said (C1-C6)alkyl, heterocycloalkyl, aryl, and heteroaryl are independently selected from the group consisting of -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, and NR'R''; each of the (C1-C6) alkyl, (C1-C6) alkenyl, (C1-C6) alkynyl, -O(C1-C6) alkyl, -O(C1-C6) alkenyl, and -O(C1-C6) alkynyl is optionally substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine; R z each occurrence of is independently H or (C1-C6) alkyl; B is [ka] is a functional group selected from D is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl, wherein the (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl is optionally substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine, or D is [ka] and Each occurrence of Z is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20)alkynyl, wherein the (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl is optionally substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine; m is 0, 1, 2, 3, 4, or 5; where n is 0, 1, 2, 3, 4, or 5).

[0133] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein m is 0, 1, 2, 3, 4, or 5.

[0134] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein m is 0, 1, 2, 3, or 4.

[0135] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein m is 0, 1, 2, or 3.

[0136] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein m is 0, 1, or 2.

[0137] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein m is 2.

[0138] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein m is 1.

[0139] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein m is 0.

[0140] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, where n is 0, 1, 2, 3, 4, or 5.

[0141] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, where n is 0, 1, 2, 3, or 4.

[0142] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, where n is 0, 1, 2, or 3.

[0143] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, where n is 0, 1, or 2.

[0144] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein n is 2.

[0145] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein n is 1.

[0146] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein n is 0.

[0147] In some embodiments, a composition of the invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' and R'' are independently H, (C1-C6) alkyl, (C1-C6) alkenyl, or (C1-C6) alkynyl, wherein the (C1-C6) alkyl, (C1-C6) alkenyl, and (C1-C6) alkynyl are optionally substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine.

[0148] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' and R'' are independently H, (C1-C6) alkyl, (C1-C6) alkenyl, or (C1-C6) alkynyl, wherein the (C1-C6) alkyl, (C1-C6) alkenyl, and (C1-C6) alkynyl are optionally substituted with 1 to 4 substituents independently selected from -OH and -O(CH3).

[0149] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' and R'' are independently H, (C1-C4) alkyl, (C1-C4) alkenyl, or (C1-C4) alkynyl, wherein the (C1-C4) alkyl, (C1-C4) alkenyl, and (C1-C4) alkynyl are optionally substituted with one or two substituents independently selected from -OH and -O(CH3).

[0150] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' and R'' are independently H and (C1-C4) alkyl, wherein the (C1-C4) alkyl is optionally substituted with one or two substituents independently selected from -OH and -O(CH3).

[0151] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' and R'' are independently H and (C1-C4) alkyl.

[0152] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' and R'', together with the nitrogen to which they are attached, are joined together to form a (C3-C6)heterocycloalkyl, wherein the (C3-C6)heterocycloalkyl is optionally substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0153] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a is H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted with 1 to 4 substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine.

[0154] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R ais H, —OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, and —O(C1-C4)alkynyl are optionally substituted by one or two substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine.

[0155] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a is H, —OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, and —O(C1-C4)alkynyl are optionally substituted by one or two substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine.

[0156] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a is H, —OH, (C1-C4) alkyl, (C1-C4) alkenyl, (C1-C4) alkynyl, —O(C1-C4) alkyl, —O(C1-C4) alkenyl, —O(C1-C4) alkynyl, chlorine, fluorine, or NR′R″.

[0157] In some embodiments, the compounds of the present invention are represented by the structure set forth in Formula I, wherein R a is NR'R''.

[0158] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a is NR'R''.

[0159] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is pentyl.

[0160] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is butyl.

[0161] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is propyl.

[0162] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is ethyl.

[0163] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is methyl.

[0164] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is H.

[0165] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a’ is H, —OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, and —O(C1-C4)alkynyl are optionally substituted by one or two substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine.

[0166] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a’ is H, —OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, and —O(C1-C4)alkynyl are optionally substituted by one or two substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine.

[0167] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a’is H, —OH, (C1-C4) alkyl, (C1-C4) alkenyl, (C1-C4) alkynyl, —O(C1-C4) alkyl, —O(C1-C4) alkenyl, —O(C1-C4) alkynyl, chlorine, fluorine, or NR′R″.

[0168] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a’ is NR'R''.

[0169] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is pentyl.

[0170] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is butyl.

[0171] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is propyl.

[0172] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is ethyl.

[0173] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is methyl.

[0174] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is H.

[0175] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a’’ is H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted by one or two substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine.

[0176] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a’’ is H, —OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, and —O(C1-C4)alkynyl are optionally substituted by one or two substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine.

[0177] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a’’is H, —OH, (C1-C4) alkyl, (C1-C4) alkenyl, (C1-C4) alkynyl, —O(C1-C4) alkyl, —O(C1-C4) alkenyl, —O(C1-C4) alkynyl, chlorine, fluorine, or NR′R″.

[0178] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a’’ is NR'R''.

[0179] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is pentyl.

[0180] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is butyl.

[0181] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is propyl.

[0182] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is ethyl.

[0183] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is methyl.

[0184] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R' is H and R'' is H.

[0185] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R a’’ is H.

[0186] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R b is H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, and —O(C1-C6)alkynyl are optionally substituted by one or two substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine.

[0187] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R b is H, —OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, fluorine, or NR′R″, wherein said (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, and —O(C1-C4)alkynyl are optionally substituted by one or two substituents independently selected from the group consisting of —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine.

[0188] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R b are independently H, —OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, —O(C1-C6)alkyl, —O(C1-C6)alkenyl, —O(C1-C6)alkynyl, chlorine, fluorine, or NR′R″.

[0189] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R b are independently H, —OH, (C1-C4)alkyl, —O(C1-C4)alkyl, chlorine, fluorine, or NH2.

[0190] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein R b are independently H and O(CH3).

[0191] In some embodiments, a composition of the invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein A is (C1-C6)alkyl or heterocycloalkyl, wherein the alkyl or heterocycloalkyl is selected from the group consisting of -OH, oxo, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, and NR'. R" may be substituted by 1 to 4 substituents independently selected from the group consisting of (C1-C6) alkyl, (C1-C6) alkenyl, (C1-C6) alkynyl, -O(C1-C6) alkyl, -O(C1-C6) alkenyl, and -O(C1-C6) alkynyl, and the (C1-C6) alkyl, (C1-C6) alkenyl, -O(C1-C4) alkynyl may be substituted by 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine.

[0192] In some embodiments, a composition of the invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein A is (C1-C4) alkyl or heterocycloalkyl, wherein the alkyl or heterocycloalkyl is selected from the group consisting of -OH, oxo, (C1-C4) alkyl, (C1-C4) alkenyl, (C1-C4) alkynyl, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, fluorine, and NR'. R" may be substituted with 1 to 4 substituents independently selected from the group consisting of (C1-C4) alkyl, (C1-C4) alkenyl, (C1-C4) alkynyl, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, and -O(C1-C4) alkynyl, and the (C1-C4) alkyl, (C1-C4) alkenyl, -O(C1-C4) alkynyl may be substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine.

[0193] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein A is heterocycloalkyl-C(O)-R z - or (C1-C4) alkyl-N(R z ) 2 - and R z Each occurrence of is independently H or (C1-C6) alkyl.

[0194] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein A is: [ka] (In the formula, R z is independently selected from H, (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, —O(C-C)alkyl, —O(C-C)alkenyl, and —O(C-C)alkynyl; X is CH or N; R d are independently selected from H, —OH, oxo, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, —OH, —O(C1-C4)alkyl, —O(C1-C4)alkenyl, —O(C1-C4)alkynyl, chlorine, and fluorine; n is 0, 1, 2, 3, or 4; p is 0, 1, 2, 3, or 4).

[0195] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein A is: [ka] (In the formula, R z is independently selected from H, (C-C) alkyl, (C-C) alkenyl, and (C-C) alkynyl; X is CH or N; R d are independently selected from —OH, (C1-C4) alkyl, —O(C1-C4) alkyl, chlorine, and fluorine; n is 0, 1, 2, or 3; p is 0, 1, or 2).

[0196] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein A is: [ka] (In the formula, R z are independently H or (C1-C6) alkyl; X is CH or N; R d are independently selected from —OH, (C1-C4) alkyl, —O(C1-C4) alkyl, chlorine, and fluorine; n is 0, 1, 2, or 3; p is 0, 1, or 2).

[0197] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein A is: [ka] (In the formula, R z are independently H or (C1-C6) alkyl; R d are independently selected from —OH, (C1-C4) alkyl, —O(C1-C4) alkyl, chlorine, and fluorine; n is 0, 1, 2, or 3; p is 0, 1, or 2).

[0198] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein A is: [ka] (In the formula, R z is independently selected from H or (C1-C6) alkyl.

[0199] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein B is: [ka] is selected from.

[0200] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein B is: [ka] is.

[0201] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein D is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, or (C6-C 20 ) alkynyl, wherein the (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, or (C6-C 20 ) the alkynyl is optionally substituted with 1 to 6 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine; or D is [ka] (Wherein Z is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )alkynyl, wherein said (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )Alkynyl is optionally substituted with 1 to 6 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0202] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein D is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, or (C6-C 20 ) alkynyl, or D is [ka] (Wherein Z is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl).

[0203] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein D is: [ka] wherein any carbon on the lipid chain may be substituted with -OH, -O(CH), chlorine, or fluorine; [ka] is cis or trans stereochemistry, X is O, C(R) 2 , or NR, R is selected from H, (C1-C4) alkyl, (C1-C4) alkenyl, (C1-C4) alkynyl, —OH, —O(C1-C4) alkyl, —O(C1-C4) alkenyl, —O(C1-C4) alkynyl, chlorine, and fluorine.

[0204] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula I, wherein D is: [ka] wherein any carbon on the lipid chain may be substituted with -OH, -O(CH), chlorine or fluorine; [ka] is cis or trans stereochemistry, X 1 -O-, -C(R) 2 -, or -NR-, each occurrence of R is independently selected from H, (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, —OH, —O(C-C)alkyl, —O(C-C)alkenyl, —O(C-C)alkynyl, chlorine, and fluorine; n is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; s is 0, 1, 2, 3, 4, 5, 6, 7, or 8; t is 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18).

[0205] In some embodiments, a composition of the invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia: [ka] (In the formula, R' and R" are independently selected from H, (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl, wherein said (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl are optionally substituted with one to four substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; or R' and R" together with the nitrogen to which they are attached, join together to form a (C3-C6)heterocycloalkyl, wherein said (C3-C6)heterocycloalkyl is optionally substituted with one to four substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; R b each occurrence of is -O(C1-C4)alkyl, wherein said -O(C1-C4)alkyl is optionally substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine; A is, [ka] is selected from Rz each occurrence of is independently H or (C1-C6) alkyl; R d each occurrence of is independently selected from —OH, (C1-C4)alkyl, —O(C1-C4)alkyl, chlorine, and fluorine; B is [ka] and D is [ka] wherein any carbon on the lipid chain may be substituted with -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, or fluorine; [ka] is cis or trans stereochemistry, X 1 -O-, -C(R) 2 -, or -NR-, each occurrence of R is independently selected from H, (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, —OH, —O(C-C)alkyl, —O(C-C)alkenyl, —O(C-C)alkynyl, chlorine, and fluorine; m is 0, 1, or 2; n is 0, 1, 2, or 3; p is 0, 1, or 2; q is 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; s is 1, 2, 3, 4, 5, 6, 7, or 8; t is 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18).

[0206] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein m is 2.

[0207] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein m is 1.

[0208] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein m is 0.

[0209] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R' and R'' are independently selected from H and (C1-C6) alkyl, wherein the alkyl is optionally substituted with 1 to 4 -OH and / or -O(CH3).

[0210] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R' and R'' are independently selected from H and (C1-C4) alkyl, wherein the alkyl is optionally substituted with 1 to 4 substituents independently selected from -OH and -O(CH3).

[0211] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R' is H and R'' is pentyl.

[0212] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R' is H and R'' is butyl.

[0213] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R' is H and R'' is propyl.

[0214] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R' is H and R'' is ethyl.

[0215] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R' is H and R'' is methyl.

[0216] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R' is H and R'' is H.

[0217] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R a’’ is H or (C1-C4) alkyl, said alkyl being optionally substituted with one or two independently selected -OH or -O(CH3).

[0218] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R a’’ is H or (C1-C4) alkyl.

[0219] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R a’’ is H.

[0220] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R b is —O(C1-C4)alkyl, wherein said —O(C1-C4)alkyl is optionally substituted with one or two independently selected —OH or —O(CH3).

[0221] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein R b is -O(CH3).

[0222] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula Ia, wherein A is: [ka] Selected from R z is independently H or (C1-C6) alkyl.

[0223] In some embodiments, a composition of the invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II: [ka] (In the formula, R 1 is (C1-C6) alkyl, wherein said (C1-C6) alkyl is optionally substituted with 1 to 4 substituents independently selected from -OH and -O(CH3), R 2 is H, methyl or -O(CH3), R 3each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or —O(C1-C4)alkyl, wherein said (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or —O(C1-C4)alkyl is optionally substituted with one or two substituents independently selected from —OH and —O(CH); R 4 each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or —O(C1-C4)alkyl, wherein said (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or —O(C1-C4)alkyl is optionally substituted with one or two substituents independently selected from —OH and —O(CH); R 5 teeth, [ka] and R 6 is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )alkynyl, wherein said (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl is optionally substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine; Each occurrence of n is 4).

[0224] In some embodiments, a composition of the invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein each occurrence of n is independently 0, 1, 2, or 3.

[0225] In some embodiments, a composition of the invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein each occurrence of n is independently 0, 1, or 2.

[0226] In some embodiments, a composition of the invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein each occurrence of n is independently 0 or 1.

[0227] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein n is absent.

[0228] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 1 is ethyl, butyl (branched or straight chain), or pentyl (branched or straight chain), wherein said ethyl, butyl, or pentyl is optionally substituted with 1 to 4 substituents independently selected from -OH, -O(CH), chlorine, and fluorine.

[0229] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 1 is butyl (branched or straight chain), said butyl being optionally substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0230] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 1 is ethyl, butyl (branched or straight chain), or pentyl (branched or straight chain), wherein said ethyl, butyl, or pentyl is optionally substituted with one or two substituents independently selected from -OH and -O(CH).

[0231] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 1 is butyl (branched or straight chain), said butyl being optionally substituted with one or two substituents independently selected from -OH and -O(CH3).

[0232] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 2 is H or methyl.

[0233] In some embodiments, the compounds of the present invention are represented by the structure set forth in Formula II, wherein R 2 is H.

[0234] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 3 each occurrence is independently a (C1-C4) alkyl, a (C1-C4) alkenyl, or —O(C1-C4) alkyl, wherein said (C1-C4) alkyl, (C1-C4) alkenyl, or —O(C1-C4) alkyl is optionally substituted with 1 to 4 substituents independently selected from —OH or —O(CH3).

[0235] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 3 each occurrence is independently (C1-C4) alkyl or —O(C1-C4) alkyl, wherein said (C1-C4) alkyl or —O(C1-C4) alkyl is optionally substituted with 1 to 4 substituents independently selected from —OH or —O(CH3).

[0236] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R3 each occurrence is independently (C1-C4) alkyl or —O(C1-C4) alkyl, wherein said (C1-C4) alkyl or —O(C1-C4) alkyl is optionally substituted with one or two substituents independently selected from —OH or —O(CH3).

[0237] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 3 Each occurrence of is independently methyl or —O(CH 3 ).

[0238] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 3 Each occurrence of is -O(CH3).

[0239] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 4 each occurrence is independently (C1-C4) alkyl or —O(C1-C4) alkyl, wherein said (C1-C4) alkyl or —O(C1-C4) alkyl is optionally substituted with 1 to 4 substituents independently selected from —OH, —O(CH3), chlorine, and fluorine.

[0240] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 4 Each occurrence of is independently (C1-C4) alkyl or —O(C1-C4) alkyl.

[0241] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 is (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl, wherein said (C6-C20 ) alkyl and (C6-C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0242] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 is (C8-C 20 ) alkyl and (C8-C 20 ) alkenyl, wherein said (C-C 20 ) alkyl and (C8-C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0243] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 is (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl, wherein (C 10 -C 20 ) alkyl and (C 10 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0244] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 is (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl, wherein (C 12 -C 20 ) alkyl and (C 12 -C 20) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0245] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 is (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl, wherein (C 14 -C 20 ) alkyl and (C 14 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0246] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 is (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl, wherein (C 16 -C 20 ) alkyl and (C 16 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0247] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 is (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl.

[0248] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 (C8-C20 ) alkyl and (C8-C 20 ) alkenyl.

[0249] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl.

[0250] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 is (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl.

[0251] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl.

[0252] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula II, wherein R 6 is (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl.

[0253] The present invention further provides a composition comprising a compound having a structure according to formula IIa: [ka] (In the formula, R 1 is butyl, said butyl being optionally substituted with one or two -OH groups; R 3 each occurrence of is independently H or —O(CH), R 5 teeth, [ka] and R 6 is (C 10 -C 20 ) alkyl, (C 10 -C 20 ) alkenyl, and (C 10 -C 20 ) alkynyl).

[0254] In some embodiments, the compounds of the present invention are represented by the structure set forth in formula IIa, wherein R 6 is (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl, wherein (C 10 -C 20 ) alkyl and (C 10 -C 20 ) The alkenyl may be optionally substituted with one or two substituents independently selected from -OH and -O(CH3).

[0255] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIa, wherein R 6 is (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl, wherein (C 12 -C 20 ) alkyl and (C 12 -C 20) The alkenyl may be optionally substituted with one or two substituents independently selected from -OH and -O(CH3).

[0256] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIa, wherein R 6 is (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl, wherein (C 14 -C 20 ) alkyl and (C 14 -C 20 ) The alkenyl may be optionally substituted with one or two substituents independently selected from -OH and -O(CH3).

[0257] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIa, wherein R 6 is (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl, wherein (C 16 -C 20 ) alkyl and (C 16 -C 20 ) The alkenyl may be optionally substituted with one or two substituents independently selected from -OH and -O(CH3).

[0258] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIa, wherein R 6 is (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl.

[0259] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIa, wherein R 6 is (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl.

[0260] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIa, wherein R 6 is (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl.

[0261] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIa, wherein R 6 is (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl.

[0262] The present invention provides compositions comprising a compound having a structure according to Formula III: [ka] (In the formula, R 1 is (C1-C6) alkyl, wherein said (C1-C6) alkyl is optionally substituted with 1 to 4 substituents independently selected from -OH and -O(CH3), R 2 is H, methyl or -O(CH3), R 3each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or —O(C1-C4)alkyl, wherein said (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or —O(C1-C4)alkyl is optionally substituted with one or two substituents independently selected from —OH and —O(CH); R 4 is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )alkynyl, wherein said (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl is optionally substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine; n is 4).

[0263] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula III, where n is 0, 1, 2, or 3.

[0264] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula III, where n is 0, 1, or 2.

[0265] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula III, where n is 0 or 1.

[0266] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula III, wherein n is absent.

[0267] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 1 is ethyl, butyl (branched or straight chain), or pentyl (branched or straight chain), wherein said ethyl, butyl, or pentyl is optionally substituted with 1 to 4 substituents independently selected from -OH, -O(CH), chlorine, and fluorine.

[0268] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 1 is butyl (branched or straight chain), said butyl being optionally substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0269] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 1 is ethyl, butyl (branched or straight chain), or pentyl (branched or straight chain), wherein said ethyl, butyl, or pentyl is optionally substituted with one or two substituents independently selected from -OH and -O(CH).

[0270] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 1 is butyl (branched or straight chain), said butyl being optionally substituted with one or two substituents independently selected from -OH and -O(CH3).

[0271] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 2 is H or methyl.

[0272] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 2 is H.

[0273] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 3 each occurrence is independently a (C1-C4) alkyl, a (C1-C4) alkenyl, or —O(C1-C4) alkyl, wherein said (C1-C4) alkyl, (C1-C4) alkenyl, or —O(C1-C4) alkyl is optionally substituted with 1 to 4 substituents independently selected from —OH or —O(CH3).

[0274] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 3 each occurrence is independently (C1-C4) alkyl or —O(C1-C4) alkyl, wherein said (C1-C4) alkyl or —O(C1-C4) alkyl is optionally substituted with 1 to 4 substituents independently selected from —OH or —O(CH3).

[0275] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 3 each occurrence is independently (C1-C4) alkyl or —O(C1-C4) alkyl, wherein said (C1-C4) alkyl or —O(C1-C4) alkyl is optionally substituted with one or two substituents independently selected from —OH or —O(CH3).

[0276] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 3 Each occurrence of is independently methyl or —O(CH 3 ).

[0277] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 3 Each occurrence of is -O(CH3).

[0278] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl, wherein said (C6-C 20 ) alkyl and (C6-C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0279] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C8-C 20 ) alkyl and (C8-C 20 ) alkenyl, wherein said (C-C 20 ) alkyl and (C8-C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0280] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl, wherein (C 10 -C 20 ) alkyl and (C 10 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0281] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl, wherein (C 12 -C 20 ) alkyl and (C 12 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0282] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl, wherein (C 14 -C 20 ) alkyl and (C 14 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0283] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl, wherein (C 16 -C 20 ) alkyl and (C 16 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0284] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl.

[0285] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C8-C 20 ) alkyl and (C8-C 20 ) alkenyl.

[0286] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl.

[0287] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl.

[0288] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl.

[0289] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula III, wherein R 4 is (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl.

[0290] The present invention also provides compositions comprising a compound having a structure according to Formula IIIa: [ka] (In the formula, R 1 is butyl, said butyl being optionally substituted with one or two -OH groups; R 3 each occurrence of is independently H or —O(CH), R 4 is (C 10 -C 20 ) alkyl, (C 10 -C 20 ) alkenyl, and (C 10 -C 20 ) alkynyl).

[0291] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl, wherein said (C6-C 20 ) alkyl and (C6-C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0292] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C8-C 20) alkyl and (C8-C 20 ) alkenyl, wherein said (C-C 20 ) alkyl and (C8-C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0293] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl, wherein (C 10 -C 20 ) alkyl and (C 10 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0294] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl, wherein (C 12 -C 20 ) alkyl and (C 12 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0295] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl, wherein (C14 -C 20 ) alkyl and (C 14 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0296] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl, wherein (C 16 -C 20 ) alkyl and (C 16 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0297] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl.

[0298] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C8-C 20 ) alkyl and (C8-C 20 ) alkenyl.

[0299] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C 10 -C 20 ) alkyl and (C 10 -C 20) alkenyl.

[0300] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl.

[0301] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl.

[0302] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in Formula IIIa, wherein R 4 is (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl.

[0303] The present invention provides compositions comprising a compound having a structure according to formula IV: [ka] (In the formula, R 1 is (C1-C6) alkyl, wherein said (C1-C6) alkyl is optionally substituted with 1 to 4 substituents independently selected from -OH and -O(CH3), R 2 is H, methyl or -O(CH3), R 3each occurrence is independently H, (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, or —O(C-C)alkyl, wherein said (C-C)alkyl, (C-C)alkenyl, (C-C)alkynyl, and —O(C-C)alkyl are optionally substituted with one or two substituents independently selected from —OH and —O(CH); R 4 Each occurrence of (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )alkynyl, wherein the (C-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) alkynyl is optionally substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, or fluorine; n is 4).

[0304] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, where n is 0, 1, 2, or 3.

[0305] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, where n is 0, 1, or 2.

[0306] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, where n is 0 or 1.

[0307] In some embodiments, compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein n is absent.

[0308] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 1 is ethyl, butyl (branched or straight chain), or pentyl (branched or straight chain), wherein said ethyl, butyl, or pentyl is optionally substituted with 1 to 4 substituents independently selected from -OH, -O(CH), chlorine, and fluorine.

[0309] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 1 is butyl (branched or straight chain), said butyl being optionally substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0310] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 1 is ethyl, butyl (branched or straight chain), or pentyl (branched or straight chain), wherein said ethyl, butyl, or pentyl is optionally substituted with one or two substituents independently selected from -OH and -O(CH).

[0311] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 1 is butyl (branched or straight chain), said butyl being optionally substituted with one or two substituents independently selected from -OH and -O(CH3).

[0312] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 2 is H or methyl.

[0313] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 2 is H.

[0314] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 3 are independently (C1-C4) alkyl, (C1-C4) alkenyl, or —O(C1-C4) alkyl, and the (C1-C4) alkyl, (C1-C4) alkenyl, or —O(C1-C4) alkyl is optionally substituted with 1 to 4 substituents independently selected from —OH or —O(CH3).

[0315] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 3 each occurrence is independently (C1-C4) alkyl or —O(C1-C4) alkyl, wherein said (C1-C4) alkyl or —O(C1-C4) alkyl is optionally substituted with 1 to 4 substituents independently selected from —OH or —O(CH3).

[0316] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 3 each occurrence is independently (C1-C4) alkyl or —O(C1-C4) alkyl, wherein said (C1-C4) alkyl or —O(C1-C4) alkyl is optionally substituted with one or two substituents independently selected from —OH or —O(CH3).

[0317] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 3 Each occurrence of is independently methyl or —O(CH 3 ).

[0318] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 3 Each occurrence of is -O(CH3).

[0319] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl, wherein the (C-C 20 ) alkyl and (C6-C 20 ) The alkenyl may be substituted with 1 to 6 independently selected -OH or -O(CH3).

[0320] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C8-C 20 ) alkyl and (C8-C 20 ) alkenyl, wherein the (C-C 20 ) alkyl and (C8-C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0321] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl, wherein (C 10 -C 20 ) alkyl and (C 10 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0322] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl, wherein (C 12 -C 20 ) alkyl and (C 12 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0323] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl, wherein (C 14 -C 20 ) alkyl and (C 14 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0324] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl, wherein (C 16 -C 20 ) alkyl and (C 16 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0325] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl.

[0326] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C8-C 20 ) alkyl and (C8-C 20 ) alkenyl.

[0327] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl.

[0328] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl.

[0329] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl.

[0330] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IV, wherein R 4 Each occurrence of (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl.

[0331] The present invention provides compositions of the invention comprising a compound having a structure according to Formula IVa: [ka] (In the formula, R 1 is butyl, said butyl being optionally substituted with one or two -OH groups; R 3 each occurrence of is independently H or —O(CH), R 4 Each occurrence of (C 10 -C 20 ) alkyl, (C 10 -C 20 ) alkenyl, and (C 10 -C 20 )alkynyl).

[0332] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl, wherein the (C-C 20 ) alkyl and (C6-C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0333] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C8-C 20 ) alkyl and (C8-C 20 ) alkenyl, wherein the (C-C 20 ) alkyl and (C8-C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0334] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl, wherein (C 10 -C 20 ) alkyl and (C 10 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0335] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl, wherein (C 12 -C 20 ) alkyl and (C 12 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0336] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl, wherein (C 14 -C 20 ) alkyl and (C 14 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0337] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl, wherein (C 16 -C 20 ) alkyl and (C 16 -C 20 ) The alkenyl may be optionally substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0338] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C6-C 20 ) alkyl and (C6-C 20 ) alkenyl.

[0339] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C8-C 20 ) alkyl and (C8-C 20 ) alkenyl.

[0340] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C 10 -C 20 ) alkyl and (C 10 -C 20 ) alkenyl.

[0341] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C 12 -C 20 ) alkyl and (C 12 -C 20 ) alkenyl.

[0342] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C 14 -C 20 ) alkyl and (C 14 -C 20 ) alkenyl.

[0343] In some embodiments, the compositions of the invention comprise one or more Streptococcus pneumoniae antigens and a compound represented by the structure set forth in formula IVa, wherein R 4 Each occurrence of (C 16 -C 20 ) alkyl and (C 16 -C 20 ) alkenyl.

[0344] In some embodiments, the present invention provides a method for the treatment of HIV-1-associated ... (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, (S)—N-(5-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, (S)-1-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)octadecan-1-one, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)tetradecanamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)oleamide, (9Z,12Z)-N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)octadeca-9,12-dienamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepan-1-yl)-5-oxopentyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperidin-1-yl)-5-oxopentyl)stearamide, N-(5-(3-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)azetidin-1-yl)-5-oxopentyl)stearamide, 1-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperidine-4-carboxamide, (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-octadecylcyclobutane-1-carboxamide, (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-hexadecylcyclobutane-1-carboxamide, N-(3-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-3-oxopropyl)stearamide, N-(7-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-7-oxoheptyl)stearamide, N-(3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)cyclobutyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-4,4-dimethyl-5-oxopentyl)stearamide, N-(6-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-methyl-6-oxohexan-2-yl)stearamide, 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecyloxy)pentan-1-one, 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecylamino)pentan-1-one, N-(5-(4-(4-((5-amino-7-(butylamino)-3-methyl-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, (9Z,12Z)-N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)octadeca-9,12-dienamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)tetradecanamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)oleamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)-4-oxobutyl)stearamide, (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)carbamate, 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperazine-1-carboxamide, 3-stearamidopropyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazine-1-carboxylate, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-6,6,6-trifluorohexyl)stearamide, N-(4-((4-((7-(butylamino)-5-hydroxy-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, and (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)carbamate, and a compound selected from the group consisting of:

[0345] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0346] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0347] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (S)—N-(5-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0348] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (S)-1-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)octadecan-1-one, or a pharmaceutically acceptable salt thereof.

[0349] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0350] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)tetradecanamide, or a pharmaceutically acceptable salt thereof.

[0351] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)oleamide, or a pharmaceutically acceptable salt thereof.

[0352] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (9Z,12Z)-N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)octadeca-9,12-dienamide, or a pharmaceutically acceptable salt thereof.

[0353] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepan-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0354] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperidin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0355] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(5-(3-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)azetidin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0356] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and 1-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperidine-4-carboxamide, or a pharmaceutically acceptable salt thereof.

[0357] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-octadecylcyclobutane-1-carboxamide, or a pharmaceutically acceptable salt thereof.

[0358] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-hexadecylcyclobutane-1-carboxamide, or a pharmaceutically acceptable salt thereof.

[0359] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(3-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-3-oxopropyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0360] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(7-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-7-oxoheptyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0361] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)cyclobutyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0362] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-4,4-dimethyl-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0363] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(6-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-methyl-6-oxohexan-2-yl)stearamide, or a pharmaceutically acceptable salt thereof.

[0364] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecyloxy)pentan-1-one, or a pharmaceutically acceptable salt thereof.

[0365] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecylamino)pentan-1-one, or a pharmaceutically acceptable salt thereof.

[0366] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-3-methyl-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0367] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (9Z,12Z)-N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)octadeca-9,12-dienamide, or a pharmaceutically acceptable salt thereof.

[0368] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)tetradecanamide, or a pharmaceutically acceptable salt thereof.

[0369] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)oleamide, or a pharmaceutically acceptable salt thereof.

[0370] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0371] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)-4-oxobutyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0372] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl (4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)carbamate, or a pharmaceutically acceptable salt thereof.

[0373] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperazine-1-carboxamide, or a pharmaceutically acceptable salt thereof.

[0374] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and 3-stearamidopropyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazine-1-carboxylate, or a pharmaceutically acceptable salt thereof.

[0375] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-6,6,6-trifluorohexyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0376] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((7-(butylamino)-5-hydroxy-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0377] In some embodiments, the present invention provides a composition comprising one or more Streptococcus pneumoniae antigens and (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)carbamate, or a pharmaceutically acceptable salt thereof.

[0378] Adjuvants Adjuvant-based approaches are being investigated to enhance vaccine immunogenicity and help address remaining unmet medical needs, particularly for populations that are more susceptible to infectious diseases (i.e., immunocompromised, elderly, or pediatric populations).

[0379] For example, although the incidence of invasive pneumococcal disease (IPD) has dramatically decreased in both children and adults following the introduction of pneumococcal conjugate vaccines, residual disease caused by persistent vaccine serotypes remains. Furthermore, serotype-specific immune responses vary, and the introduction of expanded valency PCVs has resulted in an overall decrease in serotype-specific immune responses. Adjuvant-based strategies to enhance the immunogenicity of PCVs may increase vaccine efficacy against challenging serotypes, provide more sustained immune responses, or allow for a shorter dosing schedule.

[0380] Preclinical data indicate that TLR7 / 8 agonists can increase the immunogenicity of PCV. Dowling et al. reported both accelerated and enhanced serotype-specific antibody responses, including binding antibody titers and functional opsonophagocytic killing, in neonatal and infant rhesus macaques immunized with PCV13 plus the TLR7 / 8 agonist 3M-052 compared with PCV13 alone (PCV13 containing an alum adjuvant). The addition of a TLR7 / 8 agonist to PCV13 also induced Th1-polarized CRM197-specific CD4 T cells and juvenile Streptococcus pneumoniae antigen-specific B cells in infant rhesus macaques and enhanced type II IFN and Th1-polarizing cytokine production after in vitro stimulation of human neonatal umbilical cord blood (JCI Insight 2017:e91020).

[0381] Adjuvant formulations The present disclosure provides an adjuvant formulation comprising: 1) one or more compounds of the invention, or pharmaceutically acceptable salt(s) thereof; 2) SPAN-85; 3) PS-20; and 4) squalene.

[0382] The present disclosure provides an adjuvant formulation comprising: 1) a compound of the invention, or a pharmaceutically acceptable salt thereof; 2) SPAN-85; 3) PS-20 or PS-80; and 4) squalene.

[0383] The present disclosure provides an adjuvant formulation comprising: 1) a compound of the invention, or a pharmaceutically acceptable salt thereof; 2) SPAN-85; 3) PS-20; and 4) squalene.

[0384] General method for preparing SNE formulations Generally, SNEs can be formed, for example, by first combining and mixing the components. Once mixed and blended, an aqueous buffer is added and mixed with the initial compound components to form a blended emulsion mixture. In some embodiments, the blended emulsion components are first subjected to coarse homogenization, followed by fine homogenization. The resulting formulation is then subjected to a final filtration step and stored at 4°C. The solution can contain one or more compounds, one or more sorbitan surfactants (e.g., PS-20; PS-80; SPAN-85), and one or more terpenes (e.g., squalene) in a specific molar ratio.

[0385] Alternatively, the process for preparing the SNEs of the present invention consists of four major steps: 1) solution preparation of a component mixture containing functional and non-functional components with an aqueous buffer, 2) SNE formation by split-flow mixing, 3) ultrafiltration, and 4) filtration.

[0386] Typically, the components are dissolved in ethanol before being sterile filtered to form a mixture. Some aqueous buffer is also prepared. The mixture and buffer stream are then combined using a T-tube or Y-mixer, then diluted immediately after the outlet and mixed with aqueous buffer to form the SNE intermediate. The SNE intermediate is then subjected to ultrafiltration or dialysis to concentrate the material and exchange it with a suitable buffer to remove residual ethanol. After diafiltration, a final concentration step is performed to achieve the final target concentration. The SNE bulk is then sterile filtered.

[0387] SNE preparation In some embodiments, a formulation is provided in which the SNE comprises: 1) about 0-75 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 0-30 mol % of SPAN-85; 3) about 0-30 mol % of PS-20 or PS-80; and 4) 25-85 mol % of squalene.

[0388] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 0-50 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 0-10 mol % of SPAN-85; 3) about 0-10 mol % of PS-20 or PS-80; and 4) about 50-80 mol % of squalene.

[0389] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 0-24 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-8 mol % of SPAN-85; 3) about 1-8 mol % of PS-20 or PS-80; and 4) about 60-75 mol % of squalene.

[0390] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 10-14 mol% of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-4 mol% of SPAN-85; 3) about 1-4 mol% of PS-20 or PS-80; and 4) about 50-80 mol% of squalene.

[0391] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 30-65 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 5-30 mol % of SPAN-85; 3) about 0.5-4 mol % of PS-20 or PS-80; and 4) about 10-40 mol % of squalene.

[0392] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 55-65 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 5-15 mol % of SPAN-85; 3) about 1-2.5 mol % of PS-20 or PS-80; and 4) about 25-35 mol % of squalene.

[0393] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 13-45 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 2-4 mol % of SPAN-85; 3) about 1.5-3 mol % of PS-20 or PS-80; and 4) about 50-82 mol % of squalene.

[0394] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 13-14 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-2 mol % of SPAN-85; 3) about 1-2 mol % of PS-20 or PS-80; and 4) about 79-81 mol % of squalene.

[0395] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 1-60 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-4 mol % of SPAN-85; 3) about 1-4 mol % of PS-20 or PS-80; and 4) about 32-97 mol % of squalene.

[0396] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 0-45 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-10 mol % of a non-ionic surfactant; and 3) about 50-85 mol % of squalene. In one aspect of this embodiment, the non-ionic surfactant comprises a mixture of SPAN-85 and PS-20 or PS-80.

[0397] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 0-75 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 0-30 mol % of SPAN-85; 3) about 0-30 mol % of PS-20; and 4) 25-85 mol % of squalene.

[0398] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 0-50 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 0-10 mol % of SPAN-85; 3) about 0-10 mol % of PS-20; and 4) about 50-80 mol % of squalene.

[0399] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 0-24 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-8 mol % of SPAN-85; 3) about 1-8 mol % of PS-20; and 4) about 60-75 mol % of squalene.

[0400] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 10-14 mol% of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-4 mol% of SPAN-85; 3) about 1-4 mol% of PS-20; and 4) about 50-80 mol% of squalene.

[0401] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 30-65 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 5-30 mol % of SPAN-85; 3) about 0.5-4 mol % of PS-20; and 4) about 10-40 mol % of squalene.

[0402] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 55-65 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 5-15 mol % of SPAN-85; 3) about 1-2.5 mol % of PS-20; and 4) about 25-35 mol % of squalene.

[0403] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 13-45 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 2-4 mol % of SPAN-85; 3) about 1.5-3 mol % of PS-20; and 4) about 50-82 mol % of squalene.

[0404] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 13-14 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-2 mol % of SPAN-85; 3) about 1-2 mol % of PS-20; and 4) about 79-81 mol % of squalene.

[0405] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 1-60 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-4 mol % of SPAN-85; 3) about 1-4 mol % of PS-20; and 4) about 32-97 mol % of squalene.

[0406] In some embodiments, a formulation is provided in which the SNE comprises: 1) about 0-45 mol % of a compound of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-10 mol % of a non-ionic surfactant; and 3) about 50-85 mol % of squalene. In one aspect of this embodiment, the non-ionic surfactant comprises a mixture of SPAN-85 and PS-20.

[0407] In some embodiments, the formulations provided above comprise: (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (compound A-1), (S)—N-(5-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (compound A-2), (S)-1-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)octadecan-1-one (compound A-3), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (compound B-1), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)tetradecanamide (compound B-2), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)oleamide (compound B-3), (9Z,12Z)-N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)octadeca-9,12-dienamide (compound B-4), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepan-1-yl)-5-oxopentyl)stearamide (compound B-5), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperidin-1-yl)-5-oxopentyl)stearamide (compound B-6), N-(5-(3-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)azetidin-1-yl)-5-oxopentyl)stearamide (compound B-7), 1-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperidine-4-carboxamide (compound B-8), (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-octadecylcyclobutane-1-carboxamide (compound B-9), (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-hexadecylcyclobutane-1-carboxamide (compound B-10), N-(3-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-3-oxopropyl)stearamide (compound B-11), N-(7-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-7-oxoheptyl)stearamide (compound B-12), N-(3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)cyclobutyl)stearamide (compound B-13), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-4,4-dimethyl-5-oxopentyl)stearamide (compound B-14), N-(6-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-methyl-6-oxohexan-2-yl)stearamide (compound B-15), 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecyloxy)pentan-1-one (compound B-16), 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecylamino)pentan-1-one (compound B-17), N-(5-(4-(4-((5-amino-7-(butylamino)-3-methyl-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (compound B-18), (9Z,12Z)-N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)octadeca-9,12-dienamide (compound C-1), N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)tetradecanamide (compound C-2), N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)oleamide (compound C-3), N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide (compound C-4), N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)-4-oxobutyl)stearamide (compound C-5), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)carbamate (compound D-1), 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperazine-1-carboxamide (compound D-2), 3-stearamidopropyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazine-1-carboxylate (compound D-3), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-6,6,6-trifluorohexyl)stearamide (compound D-4), N-(4-((4-((7-(butylamino)-5-hydroxy-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide (compound D-5), and (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)carbamate (compound D-6), or a pharmaceutically acceptable salt thereof.

[0408] In some embodiments of the present invention, a formulation is provided in which the SNE further comprises one or more additional components selected from a surfactant, a mixture of surfactants, a phospholipid, a terpene, a terpenoid, a triterpene, or a combination thereof.

[0409] In some embodiments of the invention, surfactants include polyoxyethylene sorbitan ester surfactants (commonly referred to as Tweens), particularly PS-20 and PS-80; copolymers of ethylene oxide (EO), propylene oxide (PO), and / or butylene oxide (BO), such as linear EO / PO block copolymers, sold under the trade name DOWFAX™; octoxynols, which may vary in the number of repeating ethoxy (oxy-1,2-ethanediyl) groups, with octoxynol-9 (Triton X-100, or t-octylphenoxypolyethoxyethanol) being of particular interest; (octylphenoxy)polyethoxyethanols (IGEPALCA-630 / NP-40); nonylphenol ethoxylates, such as the Tergitol™ NP series; polyoxyethylene fatty ethers derived from lauryl, cetyl, stearyl, and oleyl alcohols (known as Brij surfactants), such as triethylene glycol mononitrile. lauryl ether (Brij30); sorbitan trioleate (Span-85, Tween-85, or [2-[(2R,3S,4R)-4-hydroxy-3-[(Z)-octadec-9-enoyl]oxyoxolan-2-yl]-2-[(Z)-octadec-9-enoyl]oxyethyl](Z)-octadec-9-enoate) and sorbitan esters (commonly known as SPAN), such as sorbitan monolaurate.

[0410] In some embodiments of the present invention, a mixture of surfactants is used, such as a PS-20 / Span 85 mixture. A combination of a polyoxyethylene sorbitan ester, such as polyoxyethylene sorbitan monooleate (PS-80), and an octoxynol, such as t-octylphenoxypolyethoxyethanol (Triton X-100), is also suitable. Another useful combination includes laureth 9 and a polyoxyethylene sorbitan ester and / or an octoxynol.

[0411] In some embodiments of the invention, the amount of surfactant or emulsifier is: polyoxyethylene sorbitan ester (such as PS-20) 0.01 to 10 mol %, in particular about 1 to 4 mol %; octyl- or nonylphenoxypolyoxyethanol (such as Triton X-100 or other detergents in the Triton series) 0.001 to 10 mol %, in particular about 1 to 4 mol % w / v, in particular 0.01 to 0.1% w / v; polyoxyethylene ether (such as Laureth 9) 0.1 to 20 mol %, preferably 0.5 to 10 mol %, in particular 1 to 4 mol % or about 10% by weight.

[0412] In some embodiments of the invention, the phospholipid is selected from phosphatidylcholine (PC), phosphatidylethanolamine (PE), and naturally occurring phospholipids including phosphatidylglycerol (PG), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidic acid (phosphatidate) (PA), dipalmitoylphosphatidylcholine, monoacyl-phosphatidylcholine (lyso-PC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), N-acyl-PE, phosphoinositides, and phosphosphingolipids. Phospholipid derivatives include phosphatidic acid (DMPA, DPPA, DSPA), phosphatidylcholine (DDPC, DLPC, DMPC, DPPC, DSPC, DOPC, POPC, DEPC), phosphatidylglycerol (DMPG, DPPG, DSPG, POPG), phosphatidylethanolamine (DMPE, DPPE, DSPE DOPE), phosphatidylserine (DOPS). Fatty acids include C14:0, palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), linoleic acid (C18:2), linolenic acid (C18:3), and arachidonic acid (C20:4), C20:0, C22:0, and ethynyl. In certain embodiments of the invention, the phospholipid is phosphatidylserine, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), dilauroylphosphatidylcholine (DLPC), 1,2-dieicosenoyl-sn-glycero-3-phosphocholine, or 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC).

[0413] In some embodiments of the present invention, the terpene is selected from monoterpenes such as geraniol, terpenol, limonene, myrcene, linalool, or pinene. In some embodiments, the formulation comprises humulene, farnesene, or farnesol; diterpenes such as cafestol, kahweol, cembrene, or taxadiene; triterpenes such as squalene or squalane; tetraterpenes such as acyclic lycopene, monocyclic gamma-carotene, or bicyclic alpha- and beta-carotene; polyterpenes or sesquiterpenes consisting of norisoprenoids.

[0414] Pneumococcal vaccine composition Pneumococcal vaccines or compositions are well known (e.g., PNEUMOVAX® (Merck & Co., Inc., Rahway, New Jersey, USA)). Pneumococcal conjugate vaccines or compositions have been previously disclosed. See WO 2011 / 100151, WO 2019 / 139692, and WO 2020 / 131763.

[0415] Exemplary bacterial capsular polysaccharides from Streptococcus pneumoniae are serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20 (20A and 20B), 22F, 23A, 23B, 23F, 24F, 33F, 35B, 35F, or 38, among others.

[0416] General method for preparing capsule polysaccharides Bacterial capsular polysaccharides, particularly those that have been used as antigens, are suitable for use in the present invention and can be readily identified by methods for identifying immunogenic and / or antigenic polysaccharides. Exemplary bacterial capsular polysaccharides from Streptococcus pneumoniae are serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20 (20A and 20B), 22F, 23A, 23B, 23F, 24F, 33F, 35B, 35F, or 38, among others.

[0417] As used herein, de-O-acetylated serotype 15B (DeOAc15B) pneumococcal polysaccharide is substantially equivalent to serotype 15C pneumococcal polysaccharide and has substantially identical NMR spectra (data not shown). As used herein, de-O-acetylated serotype 15B pneumococcal polysaccharide and serotype 15C pneumococcal polysaccharide each have an O-acetyl content in the range of 0-5%, 0-4%, 0-3%, 0-2%, 0-1%, 0-0.5%, 0-0.1%, or no O-acetyl content. Spencer, B.L., et al. reported that pneumococcal polysaccharide 15C may be slightly O-acetylated (Spencer, B.L., et al., Clin. Vac. Immuno. (2017) 24(8):1-13). Thus, in any of the embodiments of the multivalent immunogenic compositions described herein, de-O-acetylated serotype 15B (DeOAc15B) can be used in place of serotype 15C. The process of de-O-acetylation is known in the art, for example, as described in Rajam et al., Clinical and Vaccine Immunology, 2007, 14(9):1223-1227.

[0418] Polysaccharides can be purified by known techniques. However, the present invention is not limited to polysaccharides purified from natural sources; polysaccharides may be obtained by other methods, such as total or partial synthesis. Capsular polysaccharides from Streptococcus pneumoniae can be prepared by standard techniques known to those skilled in the art. For example, polysaccharides can be isolated from bacteria and sized by known methods (see, e.g., EP 497524 and EP 497525), preferably by microfluidization achieved using a homogenizer or by chemical hydrolysis. Streptococcus pneumoniae strains corresponding to each polysaccharide serotype may be grown in a soy-based medium. Individual polysaccharides may then be purified through standard processes, including centrifugation, precipitation, and ultrafiltration. See, e.g., U.S. Patent Application Publication No. 2008 / 0286838 and U.S. Patent No. 5,847,112. The polysaccharide may be sized to reduce viscosity and / or to improve filterability and subsequent lot-to-lot consistency of the conjugated product.

[0419] Purified polysaccharides can be chemically activated to introduce functional groups capable of reacting with carrier proteins using standard techniques. Chemical activation of polysaccharides and subsequent conjugation to carrier proteins is accomplished by the procedures described in U.S. Patent Nos. 4,365,170, 4,673,574, and 4,902,506. Briefly, pneumococcal polysaccharides are reacted with a periodate-based oxidizing agent, such as sodium periodate, potassium periodate, or periodic acid, to produce reactive aldehyde groups by oxidative cleavage of adjacent hydroxyl groups. Suitable molar equivalents of periodate (e.g., sodium periodate, sodium metaperiodate, etc.) include 0.05 to 0.5 molar equivalents (molar ratio of periodate to polysaccharide repeating unit) or 0.1 to 0.5 molar equivalents. The periodate reaction can vary from 30 min to 24 h, depending on the diol conformation (e.g., acyclic diol, cis-diol, trans-diol), which controls the accessibility of the reactive hydroxyl group to sodium periodate.

[0420] The term "periodate" includes both periodate and periodic acid, and the term also includes metaperiodate (IO 4- ) and orthoperiodate (IO 6- ), including various salts of periodate (e.g., sodium periodate and potassium periodate). Capsular polysaccharides can be oxidized in the presence of metaperiodate or sodium periodate (NaIO). Additionally, capsular polysaccharides can be oxidized in the presence of orthoperiodate or periodic acid.

[0421] The purified polysaccharides can also be linked to a linker. Once activated or linked to a linker, each capsular polysaccharide can be separately conjugated to a carrier protein to form a glycoconjugate. Polysaccharide conjugates can be prepared by known conjugation techniques.

[0422] The polysaccharide can be attached to the linker to form a polysaccharide-linker intermediate in which the free end of the linker is an ester group. Thus, the linker is a linker in which at least one end is an ester group. The other end is selected so that it can react with the polysaccharide to form the polysaccharide-linker intermediate.

[0423] Polysaccharides can be attached to linkers using primary amine groups in the polysaccharide. In this case, the linker typically has ester groups at both ends. This allows attachment by reacting one of the ester groups with a primary amine group in the polysaccharide via nucleophilic acyl substitution. This reaction results in a polysaccharide-linker intermediate in which the polysaccharide is attached to the linker via an amide bond. The linker is therefore a bifunctional linker, providing one ester group for reaction with the primary amine groups in the polysaccharide and a second ester group for reaction with the primary amine groups in the carrier molecule. A typical linker is adipic acid N-hydroxysuccinimide diester (SIDEA).

[0424] Conjugation can also be performed indirectly, ie, by means of an additional linker used to derivatize the polysaccharide prior to conjugation to the linker.

[0425] Polysaccharides can be attached to additional linkers using a carbonyl group at the reducing end of the polysaccharide. This attachment involves two steps: (a1) reacting the carbonyl group with the additional linker, and (a2) reacting the free end of the additional linker with the linker. In these embodiments, the additional linker typically has primary amine groups at both ends, allowing step (a1) to be carried out by reacting one of the primary amine groups with a carbonyl group in the polysaccharide via reductive amination. A primary amine group reactive with the carbonyl group in the polysaccharide is used. A hydrazide group or a hydroxylamino group is preferred. The same primary amine group is typically present at both ends of the additional linker, allowing for the possibility of polysaccharide (Ps)-Ps bonding. This reaction results in a polysaccharide-additional linker intermediate in which the polysaccharide is attached to the additional linker via a C-N bond.

[0426] Polysaccharides can be attached to additional linkers using different groups in the polysaccharide, particularly carboxyl groups. This attachment involves two steps: (a1) reacting a group with the additional linker, and (a2) reacting the free end of the additional linker with the linker. In this case, the additional linker typically has primary amine groups at both ends, allowing step (a1) to be carried out by reacting one of the primary amine groups with a carboxyl group in the polysaccharide via EDAC activation. A primary amine group reactive with the EDAC-activated carboxyl group in the polysaccharide is used. A hydrazide group is preferred. The same primary amine group is typically present at both ends of the additional linker. This reaction results in a polysaccharide-additional linker intermediate in which the polysaccharide is attached to the additional linker via an amide bond.

[0427] Carrier proteins In a specific embodiment of the present invention, CRM197 is used as a carrier protein. CRM197 is a non-toxic variant (i.e., toxoid) of diphtheria toxin. CRM197 can be isolated from a culture of Corynebacterium diphtheria strain C7 (β197) grown in a cassamino acid and yeast extract-based medium. Furthermore, CRM197 can be recombinantly prepared according to the method described in U.S. Pat. No. 5,614,382. Typically, CRM 197 CRM197 is purified by a combination of ultrafiltration, ammonium sulfate precipitation, and ion exchange chromatography. In some embodiments, CRM197 is prepared in Pseudomonas fluorescens using Pfenex Expression Technology™ (Pfenex Inc., San Diego, CA).

[0428] Other suitable carrier proteins include additional inactivated bacterial toxins such as DT (diphtheria toxoid), TT (tetanus toxoid) or fragment C of TT, pertussis toxoid, cholera toxoid (e.g., as described in WO 2004 / 083251), E. coli LT, E. coli ST, and exotoxin A from Pseudomonas aeruginosa. Bacterial outer membrane proteins, such as outer membrane complex c (OMPC), porins, transferrin-binding proteins, pneumococcal surface protein A (PspA; see WO 02 / 091998), pneumococcal surface adhesin protein (PsaA), C5a peptidase from group A or B streptococci, or Haemophilus influenzae protein D, pneumococcal pneumolysin (Kuo et al., 1995, Infect Immunol. 63;2706-13), e.g., plies that have been detoxified in some manner, e.g., dPLY-GMBS (see WO 04 / 081515) or dPLY-formol, PhtX, including PhtA, PhtB, PhtD, PhtE, and fusions of Pht proteins, e.g., PhtDE fusions, PhtBE fusions (see WO 01 / 98334 and WO 03 / 54007), can also be used.Other proteins, such as ovalbumin, keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA), or purified protein derivative of tuberculin (PPD), PorB (from N. meningitidis), PD (Haemophilus influenzae protein D; see, for example, EP 0 594 610), or immunologically functional equivalents thereof, synthetic peptides (see EP 0 378 881 and EP 0 427 347), heat shock proteins (see WO 93 / 17712 and WO 94 / 03208), pertussis proteins (see WO 98 / 58668 and EP 0 471 177), cytokines, lymphokines, growth factors or hormones (see WO 91 / 01146), N19 protein (Baraldoi et al., 2004, Infect Immun Artificial proteins containing multiple human CD4+ T cell epitopes from antigens derived from various pathogens, such as IgG1 (see Falugi et al., 2001, Eur J Immunol 31:3816-3824), iron uptake proteins (see WO 01 / 72337), C. difficile toxin A or B (see WO 00 / 61761), and flagellin (see Ben-Yedidia et al., 1998, Immunol Lett 64:9), can also be used as carrier proteins.

[0429] When a multivalent vaccine is used, a second carrier can be used for one or more of the antigens in the multivalent vaccine. The second carrier protein is preferably a non-toxic, non-reactogenic protein that can be obtained in sufficient quantity and purity. The second carrier protein can also be conjugated or bound to an antigen, such as a pneumococcal polysaccharide, to enhance the antigen's immunogenicity. The carrier protein should be prepared according to standard conjugation procedures. Each capsular polysaccharide that is not conjugated to a first carrier protein can be conjugated to the same second carrier protein (e.g., each capsular polysaccharide molecule is conjugated to a single carrier protein). Each capsular polysaccharide that is not conjugated to a first carrier protein can be conjugated to two or more carrier proteins (each capsular polysaccharide molecule is conjugated to a single carrier protein). In such embodiments, each capsular polysaccharide of the same serotype is typically conjugated to the same carrier protein. CRM176, CRM228, CRM45 (Uchida et al., 1973, J Biol Chem 218:3838-3844); CRM9, CRM45 CRM102, CRM103, and CRM107, and Genetically Engineered Toxins by Nicholls and Youle, Eds.: Frankel, Maecel Dekker Other DT variants can be used as the second carrier protein, such as other mutations described in U.S. Pat. Nos. 4,709,017 and 4,950,740; deletions or mutations of Glu-148 to Asp, Gln, or Ser, and / or mutations of Ala158 to Gly; mutations of at least one or more residues Lys516, Lys526, Phe530, and / or Lys534; and other mutations disclosed in U.S. Pat. Nos. 5,917,017 and 6,455,673; or fragments disclosed in U.S. Pat. No. 5,843,711.

[0430] Conjugation by reductive amination Covalent attachment of polysaccharides to carrier proteins can be achieved by reductive amination, in which amine-reactive moieties on the polysaccharide are directly attached to primary amine groups (mainly lysine residues) on the protein. As is well known, reductive amination reactions proceed via a two-step mechanism. First, a Schiff base intermediate of formula R-CH=N-R' is formed by the reaction of an aldehyde group (R-CHO) on molecule 1 with a primary amine group (R'-NH2) on molecule 2. In the second step, the Schiff base is reduced to form an amino compound of formula R-CH2-NH-R'. While many reducing agents are available, in most cases, highly selective reducing agents such as sodium cyanoborohydride (NaCNBH3) are used because such agents specifically reduce only the imine functional group of the Schiff base.

[0431] Since all polysaccharides have an aldehyde function at the end of the chain (terminal aldehyde function), conjugation methods involving reductive amination of polysaccharides are very generally applicable. In the absence of other aldehyde functions in the repeating units (intrachain aldehyde functions), such methods make it possible to obtain conjugates in which a polysaccharide molecule is bound to a single molecule of carrier protein.

[0432] Typical reducing agents are cyanoborohydride salts such as sodium cyanoborohydride. A typically used imine-selective reducing agent is sodium cyanoborohydride, although other cyanoborohydride salts, including potassium cyanoborohydride, can be used. Differences in starting cyanide levels in sodium cyanoborohydride reagent lots and residual cyanide during the conjugation reaction can lead to inconsistent conjugation performance, varying conjugate size and conjugate Ps vs. CRM. 197 This can result in variable product attributes such as ratios. By controlling and / or reducing the free cyanide levels in the final reaction product, conjugation variability can be reduced.

[0433] Residual unreacted aldehydes on the polysaccharide may be reduced by the addition of a strong reducing agent, such as sodium borohydride. Generally, the use of a strong reducing agent is preferred. However, for some polysaccharides, it is preferable to avoid this step. For example, Streptococcus pneumoniae serotype 5 contains ketone groups that can readily react with strong reducing agents. In this case, it is preferable to bypass the reduction step to protect the antigenic structure of the polysaccharide.

[0434] After conjugation, the polysaccharide-protein conjugate is purified to remove excess conjugation reagent and residual free protein and free polysaccharide by one or more of the techniques known to those skilled in the art, including concentration / diafiltration, ultrafiltration, precipitation / elution, column chromatography, and depth filtration. See, for example, U.S. Patent No. 6,146,902. In one embodiment, the purification step is by ultrafiltration.

[0435] Pneumococcal conjugate compositions The present invention provides pneumococcal conjugate compositions comprising, consisting essentially of, or alternatively consisting of any of the polysaccharide serotype-conjugate combinations described herein and / or known in the art, together with a pharmaceutically acceptable carrier and an SNE as described above. The compositions may comprise, consist essentially of, or consist of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 or more different polysaccharide carrier protein conjugates, each of which contains a different capsular polysaccharide conjugated to a carrier protein (which may be one particular carrier protein or two or more different carrier proteins). In one embodiment, the capsular polysaccharide is selected from at least one or more of Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20 (20A or 20B), 22F, 23A, 23B, 23F, 24F, 33F, 35B, 35F, or 38. In another embodiment, the serotype is conjugated to the carrier protein CRM197.

[0436] In some embodiments, the polysaccharide contained in the polysaccharide carrier protein conjugate is selected from a polysaccharide selected from the group of serotypes consisting of serotypes: 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20 (20A and / or 20B), 22F, 23A, 23B, 23F, 24F, 33F, 35B, 35F, or 38. In other embodiments, the group of serotypes consists of 4, 6B, 9V, 14, 18C, 19F, and 23F. In other embodiments, the group of serotypes consists of 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. In another embodiment, the serotype group consists of: 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F, and 33F. In another embodiment, the serotype group consists of: 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, and 33F. In another embodiment, the serotype group consists of: 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In another embodiment, the serotype group consists of: 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated-15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In another embodiment, the serotype group consists of: 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In other embodiments, the serotype group consists of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.In another embodiment, the serotype group consists of: 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated-15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. In another embodiment, the serotype group consists of: 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

[0437] In some embodiments, the carrier protein contained in the polysaccharide carrier protein conjugate is selected from OMPC, PhtD, pLys, DT (diphtheria toxoid), TT (tetanus toxoid), fragment C of TT, pertussis toxoid, cholera toxoid, and CRM197. In other embodiments, the carrier protein is CRM197.

[0438] Administration / Dosage The compositions and formulations of the invention can be used to protect or treat humans susceptible to infectious diseases, such as pneumococcal infections, by administering the composition, formulation, or vaccine to a patient in need thereof via a systemic or mucosal route. In one embodiment, the invention provides a method of inducing an immune response against a Streptococcus pneumoniae capsular polysaccharide conjugate, comprising administering to a human an immunologically effective amount of an immunogenic composition of the invention. In another embodiment, the invention provides a method of vaccinating a human against pneumococcal infection, comprising administering to a human an immunologically effective amount of an immunogenic composition, formulation, or vaccine of the invention.

[0439] The optimal amount of components for a particular composition, formulation, or vaccine can be ascertained by standard studies involving observation of appropriate immune responses in subjects. For example, in another embodiment, dosages for human vaccination are determined by extrapolation from animal studies to human data. In another embodiment, dosages are determined empirically.

[0440] The methods of the present invention can be used to prevent and / or reduce the primary clinical syndromes caused by Streptococcus pneumoniae, including both invasive infections (meningitis, pneumonia, and bacteremia) and non-invasive infections (acute otitis media, sinusitis).

[0441] Administration of the compositions, formulations, or vaccines of the present invention may include one or more of injection via intramuscular, intraperitoneal, intradermal, or subcutaneous routes, or via mucosal administration to the oral / alimentary, respiratory, or genitourinary tracts. In one embodiment, intranasal administration is used for the treatment of pneumonia or otitis media (as it can more effectively prevent nasopharyngeal carriage of pneumococcus, thus attenuating the infection at its early stage).

[0442] The amount of conjugate in each vaccine (or composition or formulation) dose can be selected to induce an immunoprotective response without significant adverse effects. Such amounts can vary depending on the pneumococcal serotype. Generally, for polysaccharide-based conjugates, each dose contains 0.1 to 100 μg of each polysaccharide, particularly 0.1 to 10 μg, and more particularly 1 to 5 μg. For example, each dose can contain 100, 150, 200, 250, 300, 400, 500, or 750 ng, or 1, 1.5, 2, 3, 4, 5, 6, 7, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 20, 22, 25, 30, 40, 50, 60, 70, 80, 90, or 100 μg.

[0443] According to any of the methods of the present invention, in one embodiment, the subject is a human. In certain embodiments, the human patient is an infant (under 1 year old), a toddler (approximately 12-24 months old), or a child (approximately 2-5 years old). In other embodiments, the human patient is an elderly patient (>65 years old). The compositions of the present invention are also suitable for use in older children, adolescents, and adults (e.g., 18-45 years old or 18-65 years old).

[0444] In one embodiment of the method of the present invention, the composition, formulation, or vaccine of the present invention is administered as a single dose. In another embodiment, the composition, formulation, or vaccine is administered two, three, or four or more times, well spaced apart. For example, the composition, formulation, or vaccine may be administered at 1-, 2-, 3-, 4-, 5-, or 6-month intervals, or any combination thereof. The immunization schedule may follow that specified for pneumococcal vaccines. For example, the routine schedule for infants and young children against invasive disease caused by Streptococcus pneumoniae is 2, 4, 6, and 12-15 months of age. Thus, in a preferred embodiment, the composition, formulation, or vaccine is administered as a four-dose series at 2, 4, 6, and 12-15 months of age.

[0445] The compositions, formulations, or vaccines of the invention may also include one or more proteins from Streptococcus pneumoniae. Examples of Streptococcus pneumoniae proteins suitable for inclusion include those identified in International Patent Applications WO 02 / 083855 and WO 02 / 053761.

[0446] formulation In the following formulation embodiments, composition refers to a pharmaceutical composition and / or an immunogenic composition and / or a single-dose vaccine composition. In some embodiments, provided are compositions comprising one or more compounds described herein and one or more Streptococcus pneumoniae polysaccharides or one or more Streptococcus pneumoniae polysaccharide carrier protein conjugates. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide or Streptococcus pneumoniae polysaccharide carrier protein conjugate containing at least 7, or at least 10, or at least 13, or at least 15, or at least 20, or at least 24, or at least 27, or at least 30 or more Streptococcus pneumoniae serotypes. In some embodiments, provided are compositions comprising a compound described herein and a pneumococcal polysaccharide or a pneumococcal polysaccharide carrier protein conjugate containing 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 or more Streptococcus pneumoniae serotypes. In some embodiments, provided are compositions comprising a compound described herein and a pneumococcal polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F. In some embodiments, provided are compositions comprising a compound described herein and a pneumococcal polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F, and 33F. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, and 33F.In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 10A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 10A, 12F, 14, 15A, de-O-acetyl 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 10A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 10A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 10A, 12F, 14, 15A, de-O-acetyl 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 10A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B.In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetyl 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetyl 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.In some embodiments, provided are compositions comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

[0447] In some embodiments, the composition contains an additional 1, 2, 3, 4, or 5 S. pneumoniae polysaccharide carrier protein conjugates containing other S. pneumoniae serotypes known in the art.

[0448] In some embodiments, the composition contains an additional 1, 2, 3, 4, or 5 S. pneumoniae polysaccharide carrier protein conjugates containing other S. pneumoniae serotypes selected from groups 7C, 35F, 21, 34, 16F, 9N, 31, 17F, 23A, and 27.

[0449] In some embodiments, the composition contains one or two additional Streptococcus pneumoniae polysaccharide carrier protein conjugates containing other Streptococcus pneumoniae serotypes selected from groups 7C, 35F, 21, and 34.

[0450] In some embodiments, the composition contains one or two additional Streptococcus pneumoniae polysaccharide carrier protein conjugates containing other Streptococcus pneumoniae serotypes selected from groups 16F, 9N, 31, and 17F.

[0451] In some embodiments, the composition contains one or two additional Streptococcus pneumoniae polysaccharide carrier protein conjugates containing other Streptococcus pneumoniae serotypes selected from groups 23A and 27.

[0452] In some embodiments, the composition comprises a polysaccharide carrier protein conjugate wherein the protein is the carrier protein CRM197.

[0453] In some embodiments, a composition is provided comprising about 1 μg to about 200 mg of a compound of the invention, or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, wherein each of the conjugates is present at a concentration of about 0.01 μg to about 100 μg per 0.5 mL of vaccine formulation.

[0454] In some embodiments, a composition is provided comprising about 0.02 μg to about 40 mg of a compound of the invention, or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, wherein each of the conjugates is present at a concentration of about 0.002 μg to about 20 μg per 0.1 mL of vaccine formulation.

[0455] In some embodiments, a composition is provided comprising about 50 μg to about 2.1 mg of a compound of the invention, or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, wherein each of the conjugates is present at a concentration of about 0.002 μg to about 20 μg per 0.5 mL of vaccine formulation.

[0456] In some embodiments, a composition is provided comprising about 50 μg to about 10 mg of a compound of the invention, or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, wherein each of the conjugates is present at a concentration of about 0.002 μg to about 20 μg per 0.5 mL of vaccine formulation.

[0457] In some embodiments, a composition is provided comprising about 50 μg to about 10 mg of a compound of the invention, or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, wherein each of the conjugates is present at a concentration of about 0.002 μg to about 20 μg per 0.5 mL of vaccine formulation.

[0458] In some embodiments, a composition is provided comprising about 1 μg to about 200 mg of a compound of the invention, or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates being present at a concentration of about 0.01 μg to about 100 μg per 0.5 mL of vaccine formulation prepared as a co-lyophilized formulation.

[0459] In some embodiments, provided is a composition comprising about 1 μg to about 200 mg of a compound of the invention, or a pharmaceutically acceptable salt thereof, 1 μg to about 1 mg of aluminum in the form of APA, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of the conjugates being present at a concentration of about 0.01 μg to about 100 μg per 0.5 mL of vaccine formulation prepared as a co-lyophilized formulation.

[0460] In some embodiments, compositions are provided comprising about 0.05 μg to about 200 mg of a compound of the invention, or a pharmaceutically acceptable salt thereof, and further comprising SPAN-85, PS-80, or PS-20, and squalene, as highlighted in the various embodiments above. In some embodiments, compositions are provided comprising about 0.05 μg to about 200 mg of a compound of the invention, or a pharmaceutically acceptable salt thereof, and further comprising SPAN-85, PS-20, and squalene, as highlighted in the various embodiments above. In another embodiment, the compound is Compound A-1, A-2, or A-3. In another embodiment, the compound is Compound B-1, B-2, B-3, B-4, B-5, B-6, B-7, B-8, B-9, B-10, B-11, B-12, B-13, B-14, B-15, B-16, B-17, or B-18. In another embodiment, the compound is compound C-1, or C-2, or C-3, or C-4, or C-5. In another embodiment, the compound is compound D-1, or D-2, or D-3, or D-4, or D-5, or D-6.

[0461] The composition of the present invention can be administered subcutaneously, topically, orally, mucosally, intravenously, or intramuscularly. The composition is administered in an amount sufficient to induce a protective response. The composition can be administered by various routes, for example, orally, parenterally, subcutaneously, mucosally, or intramuscularly. The administered dose can vary depending on the patient's general condition, sex, weight, and age, as well as the administration route.

[0462] The compositions of the present invention as highlighted in the various embodiments above may be referred to as immunogenic compositions.

[0463] The compositions of the invention as highlighted in the various embodiments above may be referred to as vaccines or vaccine compositions.

[0464] In each of the above embodiments, the composition further comprises a Streptococcus pneumoniae polysaccharide carrier protein conjugate.

[0465] The present invention provides methods for treating or preventing pneumococcal disease by administering the compositions described above.

[0466] The present invention provides the use of the above compositions for treating or preventing pneumococcal disease.

[0467] All publications mentioned herein are incorporated by reference for the purpose of describing and disclosing methodology and materials that might be used in connection with the invention. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

[0468] Although preferred embodiments of the present invention have been described with reference to the accompanying drawings, it should be understood that the invention is not limited to those precise embodiments, and that various changes and modifications may be employed by those skilled in the art without departing from the scope or spirit of the present invention as defined in the appended claims.

[0469] The following examples illustrate the present invention but do not limit it.

[0470] [Example 1] General Methods for Making Compounds of Formulas I, Ia, II, IIa, III, IIIa, IV, and IVa The compounds can be prepared from known or readily prepared starting materials according to methods known to those skilled in the art of organic synthesis. Representative methods useful for making the compounds are described in the examples below. Alternative synthetic routes and analogous structures will be apparent to those skilled in the art of organic synthesis.

[0471] Those skilled in the art of organic synthesis will recognize that the synthesis of the polycyclic and / or heterocyclic cores contained in the compounds of the present invention may require protection of certain functional groups (i.e., derivatization for chemical compatibility with certain reaction conditions). Suitable protecting groups for the various functional groups of these compounds and methods for their installation and removal are well known in the field of organic chemistry. A summary of many of these methods can be found in Greene et al., Protective Groups in Organic Synthesis, Wiley-Interscience, New York, (1999).

[0472] Those skilled in the art of organic synthesis will also recognize that one route for synthesis of the polycyclic heterocyclic core of the compounds of the invention may be more desirable depending on the selection of pendant substituents.

[0473] Additionally, one skilled in the art will recognize that in some cases the order of reactions may differ from that presented herein to avoid functional group incompatibilities and adjust the synthetic route accordingly.

[0474] The preparation of polycyclic intermediates useful for making the polycyclic and / or heterocyclic cores of the compounds of the present invention is described in the literature and in compendia such as "Comprehensive Heterocyclic Chemistry," editions I, II, and III, published by Elsevier and edited by A.R. Katrittzky & R.J.K. Taylor. The manipulation of the required substitution patterns is also described in the available chemical literature summarized in compendia such as "Comprehensive Organic Chemistry," published by Elsevier and edited by D.H. R. Barton and W.D. Ollis; "Comprehensive Organic Functional Group Transformations," published by A.R. Katrittzky & R.J.K. Taylor; and "Comprehensive Organic Transformations," published by Wiley-CVH and edited by R.C. Larock.

[0475] The starting materials used and the intermediates prepared using the methods described in the examples below can be isolated and purified if desired using conventional techniques, including, but not limited to, filtration, distillation, crystallization, chromatography, etc. Such materials can be characterized using conventional means, including physical constants and spectral data.

[0476] Those skilled in the art will be aware of standard formulation techniques as described in the published literature and in textbooks such as Zheng, "Formulation and Analytical Development for Low-Dose Oral Drug Products," Wiley, 2009, ISBN.

[0477] Preparation of Compounds and Intermediates The present invention is illustrated by the following examples. All examples can utilize standard work-up and purification methods known to those skilled in the art. Unless otherwise specified, all temperatures are in °C (Celsius). Unless otherwise noted, all reactions are performed at room temperature. The synthetic methodologies presented herein are intended to illustrate applicable chemistry through the use of specific examples and are not indicative of the scope of the present disclosure.

[0478] General method Solvents, reagents and intermediates that are commercially available were used as received. Intermediates that are not commercially available were prepared as described below. 1 H NMR spectra are reported as ppm downfield from Me4Si, with the number of protons, multiplicity, and coupling constants in Hertz in parentheses. Where LC / MS data are presented, the observed parent ion is shown. Flash column chromatography was performed using prepacked normal-phase silica or bulk silica.

[0479] [Example 2] Methods for preparing intermediates of the compounds of the present invention Intermediate 1-1 Preparation of Compound Int.1-1 [ka] Methyl (7-(butylamino)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)carbamate [ka]

[0480] Step 1: Methyl (7-hydroxy-1H-pyrazolo[4,3-d]pyrimidin-5-yl)carbamate To a stirred mixture of methyl 4-amino-1H-pyrazole-5-carboxylate (2 g, 14.17 mmol) and 1,3-bis(methoxycarbonyl)-2-methyl-2-thiopseudoeuracil (2.92 g, 14.17 mmol) in MeOH (20 mL) was added AcOH (4.87 mL, 85 mmol) at ambient temperature. The resulting mixture was stirred for 16 h, and then NaOMe (25.5 g, 142 mmol) in MeOH was added dropwise. After the addition was complete, the reaction mixture was acidified to pH 1-2 with AcOH. The resulting solid was collected by filtration, washed with CHCN (200 mL), and then dried in vacuo to give the title compound. MS m / z (M+H) + : Calculated value 210.1, measured value 210.1. 1 H-NMR(400 MHz,DMSO-d6)δ 7.83(s,1H),3.67(s,3H).

[0481] Step 2: Methyl (7-(butylamino)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)carbamate To a stirred mixture of methyl (7-hydroxy-1H-pyrazolo[4,3-d]pyrimidin-5-yl)carbamate (5 g, 23.90 mmol), butan-1-amine (4.73 mL, 47.8 mmol), and DBU (3.92 mL, 26.3 mmol) in DMSO (100 mL) was added BOP (12.69 g, 28.7 mmol) at ambient temperature. The resulting mixture was warmed to 60 °C for 3 h. The resulting mixture was cooled to ambient temperature, then filtered and directly purified by reverse-phase chromatography (0-50% CH3CN / water with 0.1% formic acid modifier) ​​to provide the title compound. MS m / z (M+H) + : Calculated value 265.1, measured value 265.1. 1 H-NMR(400 MHz,DMSO-d6)δ 12.94(br.,1H),9.89(s,1H),8.14-7.99(m,2H),3.67(s,3H),3.56-3.51(m,2H),1.66-1.58(m,2H),1.42-1.33(m,2H),0.92(t,J=7.2 Hz,3H).

[0482] Intermediate 2-1 Preparation of Compound Int.2-1 [ka] 5-Azido-N-butyl-1H-pyrazolo[4,3-d]pyrimidin-7-amine [ka]

[0483] Step 1: N-butyl-5-chloro-1H-pyrazolo[4,3-d]pyrimidin-7-amine To a mixture of 5,7-dichloro-1H-pyrazolo[4,3-d]pyrimidine (3 g, 15.87 mmol) in THF (30 mL) was added DIEA (3.08 g, 23.81 mmol) and butan-1-amine (1.741 g, 23.81 mmol) at 0 °C. After the addition was complete, the mixture was warmed to ambient temperature and stirred for 2 h. The resulting mixture was diluted with water (200 mL) and then extracted with EtOAc (3 × 300 mL). The combined organic extracts were washed with brine (2 × 300 mL), then dried (Na SO ), and then filtered. The filtrate was concentrated to give the title compound, which was used directly in the next step without purification. MS m / z (M+H) + : Calculated value 226.1, measured value 226.2.

[0484] Step 2: 5-Azido-N-butyl-1H-pyrazolo[4,3-d]pyrimidin-7-amine To a mixture of N-butyl-5-chloro-1H-pyrazolo[4,3-d]pyrimidin-7-amine (3.5 g, 15.51 mmol) in AcOH (6 mL) and EtOH (24 mL) at ambient temperature was added sodium azide (1.512 g, 23.26 mmol), and the mixture was then heated to 100 °C for 3 h. The resulting mixture was cooled to ambient temperature, diluted with water (200 mL), and extracted with EtOAc (3 × 300 mL). The combined organic extracts were washed with brine (2 × 300 mL), dried (Na SO ), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-20% EtOAc / petroleum ether) to give the title compound. MS m / z (M+H) + : Calculated value 233.1, measured value 233.1.

[0485] Utilizing the procedure described for intermediate 2-1 and substituting the appropriate reagent for butan-1-amine, the following compounds were prepared.

[0486] [Table 2]

[0487] Intermediate 2-3 Preparation of Compound Int.2-3 [ka] 5-Azido-N-butyl-3-methyl-1H-pyrazolo[4,3-d]pyrimidin-7-amine [ka]

[0488] Step 1: 5-Azido-N-butyl-3-iodo-1H-pyrazolo[4,3-d]pyrimidin-7-amine To a solution of 5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidin-7-amine (1 g, 4.31 mmol) in DMF (30 mL) was added N-iodosuccinimide (1.937 g, 8.61 mmol), and the mixture was then heated to 40° C. After heating overnight, the mixture was cooled to room temperature and then diluted with 10% NaSO (40 mL). The resulting solid was collected by filtration, washed with water (3×10 mL), and then dried under vacuum to give the title compound. MS m / z (M+H) + :Calculated value 359.0, actual value 359.0.

[0489] Step 2: 5-Azido-N-butyl-3-methyl-1H-pyrazolo[4,3-d]pyrimidin-7-amine To a solution of 5-azido-N-butyl-3-iodo-2H-pyrazolo[4,3-d]pyrimidin-7-amine (1 g, 2.79 mmol) in 1,4-dioxane (10 mL) and HO (1 mL) was added Pd(dppf)Cl (0.204 g, 0.279 mmol), KPO (1.185 g, 5.58 mmol), and 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (3.51 g, 13.96 mmol) under N, and the mixture was heated to 100 °C overnight. The resulting mixture was cooled to room temperature, diluted with EtOAc (30 mL), dried (NaSO), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (30-70% EtOAc / petroleum ether) to give the title compound. MS m / z (M+H) + : Calculated value 247.1, actual value 247.4.

[0490] Intermediate 3-1 Preparation of Compound Int.3-1 [ka] tert-Butyl 4-(4-(hydroxymethyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate [ka]

[0491] Step 1: tert-butyl 4-(4-formyl-3,5-dimethoxyphenyl)piperazine-1-carboxylate To a stirred mixture of 4-bromo-2,6-dimethoxybenzaldehyde (5 g, 20.40 mmol) in toluene (20 mL) under an argon atmosphere, tert-butyl piperazine-1-carboxylate (5.70 g, 30.6 mmol), RuPhos (1.904 g, 4.08 mmol), Pd(dba) (1.868 g, 2.040 mmol), and NaOtBu (5.88 g, 61.2 mmol) were added, and the mixture was then heated to 100 °C. After 6 h, the mixture was cooled to ambient temperature, diluted with water (50 mL), and extracted with EtOAc (3 × 80 mL). The combined organic extracts were washed with brine (3 × 50 mL), dried (NaSO), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-100% EtOAc / DCM) to give the title compound. MS m / z (M+H) + : Calculated value 351.2, measured value 351.3.

[0492] Step 2: tert-butyl 4-(4-(hydroxymethyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate To a stirred mixture of tert-butyl 4-(4-formyl-3,5-dimethoxyphenyl)piperazine-1-carboxylate (3 g, 8.56 mmol) in EtOH (10 mL) was added NaBH (0.389 g, 10.27 mmol) at ambient temperature. After 15 min, the mixture was diluted with water (15 mL) and then extracted with EtO (3 x 50 mL). The combined organic extracts were washed with brine (3 x 50 mL), then dried (NaSO), and then filtered. The filtrate was then concentrated to give the title compound, which was used directly in the next step without purification. MS m / z (M+H) + : Calculated value 353.2, measured value 353.3.

[0493] Intermediate 4-1 Preparation of Compound Int.4-1 [ka] tert-Butyl (4-(chloromethyl)-3,5-dimethoxybenzyl)(methyl)carbamate [ka]

[0494] Step 1: tert-butyl (4-formyl-3,5-dimethoxybenzyl)(methyl)carbamate 4-Bromo-2,6-dimethoxybenzaldehyde (6 g, 24.48 mmol), N-(tert-butoxycarbonyl)-N-methylglycine (9.26 g, 49.0 mmol), Ir[dF(CF3)ppy]2(dtbbpy)PF6 (0.275 g, 0.245 mmol), and [Ni(dtbbpy)(HO)4]Cl2 (0.575 g, 1.224 mmol) were combined in DMSO (120 mL). To this was added BTMG (10 mL, 49.0 mmol). The mixture was sonicated until all solids dissolved. The resulting mixture was evenly divided into four screw-cap vials equipped with stir bars. N2 was bubbled through each mixture for 5 min. The vial was capped, and the mixture was then irradiated overnight in a PennOC Photoreactor® (wavelength: 420 nm; LED intensity: 100%; fan speed: 5000 rpm; stirring: 1200 rpm). The separated mixtures were combined, diluted with HO, and extracted with EtOAc (3x). The combined organic extracts were washed with HO and brine, then dried (MgSO), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-50% 3:1 EtOAc:EtOH / heptane) to provide the title compound. 1 H-NMR(400 MHz,CDCl3)δ 10.47(s,1H),6.44(bs,2H),4.41(s,2H),3.88(s,6H),2.87(bs,3H),1.49(bs,9H).

[0495] Step 2: tert-butyl (4-(hydroxymethyl)-3,5-dimethoxybenzyl)(methyl)carbamate A solution of tert-butyl (4-formyl-3,5-dimethoxybenzyl)(methyl)carbamate (6.98 g, 22.56 mmol) in MeOH (100 mL) was cooled to 0 °C. To this was added NaBH (1.03 g, 27.2 mmol) in portions. After 1 h, gas evolution ceased. The cooling bath was removed, and the mixture was allowed to warm to ambient temperature. After 90 min, the mixture was concentrated. The residue was taken up in DCM and filtered through a pad of Celite®, washing with DCM, and the filtrate was then concentrated. The crude product was subjected to silica gel chromatography (0-15% 3:1 EtOAc:EtOH / heptane) to give the title compound. 1 H-NMR(400 MHz,CDCl3)δ 6.45(bs,2H),4.76(d,J=6.6 Hz,2H),4.39(s,2H),3.83(s,6H),2.86(bs,3H),1.49(s,9H).

[0496] Step 3: tert-butyl (4-(chloromethyl)-3,5-dimethoxybenzyl)(methyl)carbamate A solution of N-chlorosuccinimide (0.472 g, 3.53 mmol) in DCM (1 mL) was cooled to 0 °C. To this, dimethyl sulfide (0.239 g, 3.85 mmol) was added slowly, resulting in a precipitate. The reaction mixture was cooled to -20 °C, and then a solution of tert-butyl (4-(hydroxymethyl)-3,5-dimethoxybenzyl)(methyl)carbamate (1 g, 3.21 mmol) in DCM was added dropwise. The mixture was stirred for 2 h, allowing the temperature to reach 0 °C, during which time all the solid precipitate dissolved to give a clear solution. The solution was poured into cold brine and then extracted with EtO (2x). The combined organic extracts were washed with cold brine, then dried (NaSO), then filtered, and the filtrate was concentrated to give the title compound, which was used directly in the next step without purification. MS m / z (M+H) + : Calculated value 330.1, measured value 329.2.

[0497] Intermediate 5-1 Preparation of Compound Int.5-1 [ka] tert-Butyl (4-oxobutyl)carbamate [ka] To a mixture of tert-butyl (4-hydroxybutyl)carbamate (1 g, 5.28 mmol) in CHCN (30 mL) at ambient temperature was added IBX (1.231 g, 6.34 mmol), and the mixture was then heated to 80 °C. After 1 h, the mixture was cooled to ambient temperature, diluted with water (100 mL), and extracted with EtOAc (3 × 200 mL). The combined organic extracts were washed with brine (2 × 200 mL), dried (NaSO), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0–70% EtOAc / petroleum ether) to provide the title compound. 1 H-NMR (400 MHz, CDCl3): δ 3.76-3.73(m,2H),2.53-2.49(t,2H),2.03-1.97(m,2H),1.53-1.45(s,9H).

[0498] Intermediate 6-1 Preparation of Compound Int.6-1 [ka] (6Z,9Z,28Z,31Z)-Heptatriaconta-6,9,28,31-tetraen-19-yl(4-nitrophenyl)carbonate [ka] The mixture of (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-ol (300 mg, 0.567 mmol) and hypochlorous acid 4-nitrobenzoic anhydride (229 mg, 1.134 mmol) in DCM (4.5 mL) and pyridine (1.5 mL) was stirred at ambient temperature for 5 hours.The resulting mixture was concentrated and then purified by preparative TLC (petroleum ether: DCM = 1: 1) to obtain the title compound.

[0499] Utilizing the procedure described for intermediate 6-1 and substituting the appropriate reagents for (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-ol, the following compounds were prepared:

[0500] [ka]

[0501] [Table 3]

[0502] Intermediate 7-1 Preparation of Compound Int.7-1 [ka] 4-Stearamidobutanoic acid [ka] A mixture of 4-aminobutanoic acid (1 g, 9.70 mmol) and Na2CO3 (1.542 g, 14.55 mmol) in THF (30 mL) and water (30 mL) was cooled to 0 °C. To this was added stearoyl chloride (2.94 g, 9.70 mmol). After the addition was complete, the mixture was stirred at 25 °C for 16 h. The resulting mixture was quenched with saturated KHSO4 and then extracted with CHCl3 (500 mL). The combined organic layers were washed with brine (3 x 100 mL), then dried (Na2SO4), then filtered, and the filtrate was concentrated to give the title compound. MS m / z (M−H) - : Calculated value 368.6, actual value 368.3.

[0503] Utilizing the procedure described for intermediate 7-1 and substituting the appropriate reagent for 4-aminobutanoic acid, the following compounds were prepared.

[0504] [ka]

[0505] [Table 4]

[0506] Intermediate 8-1 Preparation of Compound Int.8-1 [ka] N-(3-aminopropyl)stearamide [ka] A mixture of propane-1,3-diamine (1 g, 13.49 mmol) and pyridine (1.091 mL, 13.49 mmol) in DMF (15 mL) was cooled to 0° C. To this was added stearoyl chloride (4.09 g, 13.49 mmol). After the addition was complete, the mixture was stirred at 25° C. for 16 hours. The resulting mixture was diluted with ice water to give a solid. The solid was collected by filtration, then washed with water (3×100 mL), and then dried to give the title compound. MS m / z (M+H) + : Calculated value 341.6, measured value 341.3.

[0507] Utilizing the procedure described in Intermediate 8-1 and substituting the appropriate reagents for propane-1,3-diamine, the following compounds were prepared:

[0508] [ka]

[0509] [Table 5]

[0510] Intermediate 9-1 Preparation of Compound Int.9-1 [ka] 2-((1s,3s)-3-(octadecylcarbamoyl)cyclobutyl)acetic acid [ka]

[0511] Step 1: tert-Butyl 2-((1s,3s)-3-(octadecylcarbamoyl)cyclobutyl)acetate To a stirred mixture of (1s,3s)-3-(2-(tert-butoxy)-2-oxoethyl)cyclobutane-1-carboxylic acid (0.5 g, 2.334 mmol), HATU (1.78 g, 4.67 mmol), and DIEA (1.22 mL, 7.00 mmol) in DMF (10 mL) was added a solution of octadecan-1-amine (0.818 g, 3.03 mmol) in DMF (10 mL) at 25° C. After 12 h, the mixture was diluted with water (10 mL) and then extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (3 x 10 mL), then dried (Na2SO4), then filtered, and the filtrate was concentrated to give the title compound as an 80:20 mixture with tert-butyl 2-((1s,3s)-3-(hexadecylcarbamoyl)cyclobutyl)acetate. MS m / z (M+H) + : Calculated value 466.4, actual value 466.2.

[0512] Step 2: 2-((1s,3s)-3-(octadecylcarbamoyl)cyclobutyl)acetic acid To a stirred mixture of tert-butyl 2-(3-(octadecylcarbamoyl)cyclobutyl)acetate (0.932 g, 2 mmol) in 1,4-dioxane (16 mL) at 25° C. was added 4 M HCl in 1,4-dioxane (4 mL). After 6 h, the mixture was diluted with EtOAc and then washed with brine (3×30 mL). The organic layer was dried (NaSO) and then filtered, and the filtrate was concentrated to give the title compound as an 80:20 mixture with 2-((1s,3s)-3-(hexadecylcarbamoyl)cyclobutyl)acetic acid. MS m / z (M+H) + : Calculated value 410.4, measured value 410.4.

[0513] Intermediate 10-1 Preparation of Compound Int.10-1 [ka] 2,2-Dimethyl-5-stearamidopentanoic acid [ka] To a mixture of N-(5-hydroxy-4,4-dimethylpentyl)stearamide (Int. 8-3, 500 mg, 1.257 mmol) in acetone (10 mL) was added a solution of chromium trioxide in sulfuric acid (0.691 mL, 1.383 mmol) at 0° C. After 6 h, the mixture was diluted with water. The resulting solid was collected by filtration, washed with water, and then air-dried. MS m / z (M+H) + : Calculated value 412.4, measured value 412.1.

[0514] Intermediate 11-1 Preparation of Compound Int.11-1 [ka] 5-Methyl-5-stearamidohexanoic acid [ka]

[0515] Step 1: tert-Butyl methyl glutarate A solution of 5-(tert-butoxy)-5-oxopentanoic acid (5 g, 26.6 mmol) in DCM (40 mL) and MeOH (40 mL) was cooled to 0 °C. To this was added (diazomethyl)trimethylsilane (39.8 mL, 80 mmol, 2 M in hexanes). After the addition was complete, the mixture was allowed to warm to room temperature. After 3 h, the mixture was concentrated. The crude product was subjected to silica gel chromatography (0-5% EtOAc / petroleum ether) to give the title compound. 1H NMR(400 MHz,CDCl3)δ 3.68(s,3H),2.37(t,J=7.2 Hz,2H),2.28(t,J=7.6 Hz,2H),1.91(quin,J=7.2 Hz,2H),1.44(s,9H).

[0516] Step 2: tert-Butyl 5-hydroxy-5-methylhexanoate To a solution of tert-butyl methyl glutarate (1 g, 4.94 mmol) in THF (16 mL) was added MeMgBr (4.12 mL, 12.36 mmol, 3 M in THF) at 0 °C. After 2 h, the mixture was quenched with saturated NH4Cl (25 mL), then warmed to room temperature, and then extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (30 mL), then dried (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-20% EtOAc / petroleum ether) to give the title compound. 1 H NMR(400 MHz,CDCl3)δ 2.25(t,J=7.2 Hz,2H),1.69-1.64(m,2H),1.50-1.48(m,2H),1.45(s,9H),1.23(s,6H).

[0517] Step 3: 5-methyl-5-stearamidohexanoic acid To a solution of tert-butyl 5-hydroxy-5-methylhexanoate (300 mg, 1.483 mmol) in AcOH (2 mL) was added stearonitrile (787 mg, 2.97 mmol) at room temperature, and the mixture was then cooled to 0° C. H2SO4 (0.5 mL) was added. After the addition was complete, the cooling bath was removed and the mixture was allowed to warm to room temperature. After stirring overnight, the mixture was quenched with saturated NaHCO3 to pH=5. The resulting mixture was extracted with EtOAc (2 mL x 3). The combined organic layers were dried (Na2SO4) and then filtered, and the filtrate was concentrated. The residue was taken up in MeCN (5 mL). The resulting precipitated solid was collected and then recrystallized from EtOAc to give the title compound. MS m / z (M+H) + : Calculated value 412.7, measured value 412.3.

[0518] Intermediate 12-1 Preparation of Compound Int.12-1 [ka] 5-((tert-butoxycarbonyl)(octadecyl)amino)pentanoic acid [ka]

[0519] Step 1: Methyl 5-((tert-butoxycarbonyl)amino)pentanoate To a solution of 5-((tert-butoxycarbonyl)amino)pentanoic acid (2 g, 9.21 mmol) in MeOH (15 mL) and DCM (15 mL) was added 2 M (trimethylsilyl)diazomethane in hexanes (23.01 mL, 46.0 mmol) at room temperature. After 1 h, the mixture was concentrated to give the title compound, which was used without purification. 1 H NMR(400 MHz,CDCl3)δ 4.56(s,1H),3.68(s,3H),3.13(q,J=6.4 Hz,2H),2.34(t,J=7.2 Hz,2H),1.70-1.62(m,2H),1.49-1.56(m,2H),1.45(s,9H).

[0520] Step 2: Methyl 5-((tert-butoxycarbonyl)(octadecyl)amino)pentanoate To a solution of methyl 5-((tert-butoxycarbonyl)amino)pentanoate (1 g, 4.32 mmol) in DMF (20 mL) was added NaH (0.259 g, 6.49 mmol, 60% in mineral oil) at 0 °C. After the addition was complete, the mixture was warmed to room temperature. After 30 min, 1-bromooctadecane (4.32 g, 12.97 mmol) was added, and the mixture was then heated to 50 °C. After 2 h, the mixture was cooled to room temperature, diluted with water (100 mL), and extracted with EtOAc (3 ×). The combined organic layers were washed with brine (50 mL), dried (Na SO ), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-20% EtOAc / petroleum ether) to provide the title compound. 1H NMR(400 MHz,CD3OD)δ 3.67-3.64(m,3H),3.23-3.15(m,4H),2.39-2.33(m,2H),1.57(d,J=4.4 Hz,6H),1.46(s,9H),1.29(s,30H),0.92-0.88(m,3H).

[0521] Step 3: 5-((tert-butoxycarbonyl)(octadecyl)amino)pentanoic acid To a stirred solution of methyl 5-((tert-butoxycarbonyl)(octadecyl)amino)pentanoate (240 mg, 0.496 mmol) in THF (6 mL) and HO (2 mL) was added LiOH monohydrate (62.5 mg, 1.488 mmol) at room temperature, and the mixture was then warmed to 40° C. After stirring overnight, the mixture was cooled to room temperature and the pH was adjusted to 7 with 1 N HCl. The resulting mixture was extracted with EtOAc (3×5 mL). The combined organic layers were washed with brine (3×5 mL), dried (NaSO), filtered, and the filtrate was concentrated to give the title compound, which was used in the next step without purification. 1 H NMR(400 MHz,CD3OD)δ 3.23-3.14(m,4H),2.35-2.28(m,2H),1.65-1.52(m,6H),1.46(s,9H),1.29(s,30H),0.92-0.88 ppm(m,3H).

[0522] Intermediate 13-1 Preparation of Compound Int.13-1 [ka] 5-(octadecyloxy)pentanoic acid [ka]

[0523] Step 1: 1-(hex-5-en-1-yloxy)octadecane To a solution of octadecan-1-ol (5 g, 18.48 mmol) in DMF (100 mL) was added NaH (3.70 g, 92 mmol) (60% in mineral oil) in portions at room temperature. After stirring for 30 minutes, 6-bromo-1-hexene (24.70 mL, 185 mmol) was added, and the mixture was then heated to 80 °C. After heating overnight, the mixture was cooled to room temperature, diluted with water (200 mL), and extracted with EtOAc (100 mL × 3). The combined organic layers were dried (Na2SO4), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-100% EtOAc / petroleum ether) to give the title compound. 1 H NMR(400 MHz,CDCl3)δ 5.82(ddt,J=17.2,10.4,6.8,6.8 Hz,1H),5.01(dq,J=17.6,1.2 Hz,1H),4.95(d,J=10.0 Hz,1H),3.45-3.36(m,4H),2.08(d,J=7.2 Hz,2H),1.65-1.55(m,5H),1.49-1.44(m,2H),1.39-1.1.17(m,29H),0.85-0.92(m,3H).

[0524] Step 2: 5-(Octadecyloxy)pentanoic acid To a solution of 1-(hex-5-en-1-yloxy)octadecane (1 g, 2.84 mmol) in MeCN (10 mL) was added a solution of RuCl (0.059 g, 0.284 mmol) and NaIO (2.426 g, 11.34 mmol) in HO (10 mL) at room temperature. After 2 hours, the mixture was diluted with NaSO (25 mL) and extracted with EtOAc (25 mL × 3). The combined organic layers were washed with 1N HCl (20 mL), saturated NaHCO (20 mL), and brine (20 mL), then dried (NaSO), filtered, and the filtrate was concentrated to give the title compound, which was used in the next step without purification. 1H NMR(400 MHz,CDCl3)δ ppm 3.39-3.44(m,4 H),2.40(t,J=7.27 Hz,2 H),1.70-1.75(m,2 H),1.65(br d,J=7.51 Hz,2 H),1.55-1.58(m,2 H),1.26(s,30 H),0.87-0.89(m,3 H).

[0525] Intermediate 14-1 Preparation of Compound Int.14-1 [ka] 2-(4-(1,4-diazepan-1-yl)-2-methoxybenzyl)-5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidin-7-amine [ka]

[0526] Step 1: tert-butyl 1-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperidine-4-carboxylate To a mixture of 5-azido-2-(4-bromo-2-methoxybenzyl)-N-butyl-2H-pyrazolo[4,3-d]pyrimidin-7-amine (from compound B-1, Step 1, 500 mg, 1.159 mmol) and tert-butyl 1,4-diazepane-1-carboxylate (302 mg, 1.507 mmol) in toluene (10 mL) was added RuPhos (216 mg, 0.464 mmol), Pd(dba) (212 mg, 0.232 mmol), and CsCO (1133 mg, 3.48 mmol) under N, and the mixture was then heated to 110 °C. After heating overnight, the mixture was cooled to room temperature and then diluted with water (100 mL), and the resulting mixture was then extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (3 x 300 mL), then dried (NaSO), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-52% 3:1 EtOAc / petroleum ether) to give the title compound. MS m / z (M+H) + : Calculated value 551.7, measured value 551.1.

[0527] Step 2: 2-(4-(1,4-diazepan-1-yl)-2-methoxybenzyl)-5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidin-7-amine A solution of tert-butyl 4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepane-1-carboxylate (350 mg, 0.636 mmol) in DCE (2.00 mL) was cooled to 0° C. To this was added 4 M HCl in 1,4-dioxane (2 mL) at 0° C. After 1 h, the mixture was warmed to room temperature and then concentrated to give the title compound as the HCl salt, which was used without purification. MS m / z (M+H) + : Calculated value 451.3, measured value 451.3.

[0528] Utilizing the procedure described for intermediate 14-1 and substituting the appropriate reagent for tert-butyl 1,4-diazepane-1-carboxylate, the following compounds were prepared.

[0529] [ka]

[0530] [Table 6]

[0531] Intermediate 15-1 Preparation of Compound Int.15-1 [ka] 5-Azido-N-butyl-2-(2-methoxy-4-(piperidin-4-yl)benzyl)-2H-pyrazolo[4,3-d]pyrimidin-7-amine [ka]

[0532] Step 1: tert-butyl 4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-3,6-dihydropyridine-1(2H)-carboxylate To a mixture of 5-azido-2-(4-bromo-2-methoxybenzyl)-N-butyl-2H-pyrazolo[4,3-d]pyrimidin-7-amine (derived from compound B-1, Step 1, 400 mg, 0.927 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (430 mg, 1.391 mmol) in 1,4-dioxane (4 mL) / HO (0.4 mL) was added CsCO (604 mg, 1.855 mmol) and X-PhosPdG (39.3 mg, 0.046 mmol) under N, and the mixture was then heated to 80 °C. After 2 h, the mixture was cooled to room temperature and then diluted with EtOAc (200 mL). The resulting mixture was washed with brine (3 x 100 mL), then dried (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-50% EtOAc / petroleum ether) to give the title compound. MS m / z (M+H) + : Calculated value 534.3, measured value 534.4.

[0533] Step 2: tert-butyl 4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperidine-1-carboxylate To a solution of tert-butyl 4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-3,6-dihydropyridine-1(2H)-carboxylate (290 mg, 0.543 mmol) in MeOH (10 mL) was added Pd / C (150 mg, 1.410 mmol) under N. The resulting mixture was placed under and atmosphere of H (3 x vacuum / H) and stirred at room temperature. After 16 h, the mixture was degassed (3 x vacuum / N), then filtered, washing with MeOH, and the filtrate was concentrated to give the title compound. MS m / z (M+H) + : Calculated value 536.3, measured value 536.3.

[0534] Step 3: 5-Azido-N-butyl-2-(2-methoxy-4-(piperidin-4-yl)benzyl)-2H-pyrazolo[4,3-d]pyrimidin-7-amine To a solution of tert-butyl 4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperidine-1-carboxylate (250 mg, 0.467 mmol) in DCM (2 mL) was added 4 M HCl in 1,4-dioxane (2 mL) at room temperature. After 1 h, the mixture was concentrated to give the title compound as the HCl salt, which was used without purification. MS m / z (M+H) + : Calculated value 436.3, measured value 436.6.

[0535] Intermediate 16-1 Preparation of Compound Int.16-1 [ka] 2-(4-(azetidin-3-yl)-2-methoxybenzyl)-5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidin-7-amine [ka]

[0536] Step 1: tert-butyl 3-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)azetidine-1-carboxylate To a solution of NiI (36.2 mg, 0.116 mmol) in DMA (1 mL) was added picolinimidamide hydrochloride (36.5 mg, 0.232 mmol), and the mixture was then heated to 50 °C. After 30 min, zinc powder (152 mg, 2.319 mmol) was added, followed by a solution of 5-azido-2-(4-bromo-2-methoxybenzyl)-N-butyl-2H-pyrazolo[4,3-d]pyrimidin-7-amine (derived from compound B-1, step 1, 500 mg, 1.159 mmol) and tert-butyl 3-bromoazetidine-1-carboxylate (411 mg, 1.739 mmol) in DMA (10 mL). After stirring overnight at room temperature, the mixture was directly purified by reverse-phase chromatography (CHCN / water with 0.1% TFA modifier) ​​to give the title compound. MS m / z (M+H) + : Calculated value 508.3, measured value 508.4.

[0537] Step 2: 2-(4-(azetidin-3-yl)-2-methoxybenzyl)-5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidin-7-amine To a solution of tert-butyl 3-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)azetidine-1-carboxylate (60 mg, 0.118 mmol) in DCM (0.5 mL) was added 4 M HCl in 1,4-dioxane (0.5 mL) at room temperature. After 1 h, the mixture was concentrated to give the title compound as the HCl salt, which was used without purification. MS m / z (M+H) + : Calculated value 408.2, measured value 408.3.

[0538] Intermediate 17-1 Preparation of Compound Int.17-1 [ka] (S)-5-Azido-N-(1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)-2-(2,6-dimethoxy-4-(piperazin-1-yl)benzyl)-2H-pyrazolo[4,3-d]pyrimidin-7-amine [ka]

[0539] Step 1: (4-bromo-2,6-dimethoxyphenyl)methanol To a solution of 4-bromo-2,6-dimethoxybenzaldehyde (5 g, 20.40 mmol) in EtOH (60 mL) was added NaBH (0.926 g, 24.48 mmol) at room temperature. After 1 h, the mixture was diluted with water (50 mL) and then extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine (50 mL), then dried (NaSO), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (15-50% EtOAc / petroleum ether) to give the title compound. 1H NMR(400 MHz,CDCl3)δ 6.72(s,2H),4.72(d,J=6.8 Hz,2H),3.84(s,6H),2.32(t,J=6.8 Hz,1H).

[0540] Step 2: 5-Bromo-2-(chloromethyl)-1,3-dimethoxybenzene To a solution of (4-bromo-2,6-dimethoxyphenyl)methanol (1 g, 4.05 mmol) in DCM (10 mL) was added SOCl (0.591 mL, 8.09 mmol) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred overnight. The resulting mixture was concentrated to give the title compound, which was used in the next step without purification. 1 H NMR(400 MHz,CDCl3)δ 6.72(s,2H),4.70(s,2H),3.87(s,6H).

[0541] Step 3: (S)-2-((5-azido-2-(4-bromo-2,6-dimethoxybenzyl)-2H-pyrazolo[4,3-d]pyrimidin-7-yl)amino)pentan-1-ol To a solution of (S)-2-((5-azido-2H-pyrazolo[4,3-d]pyrimidin-7-yl)amino)pentan-1-ol (Int. 2-2, 800 mg, 3.05 mmol) in DMF (15 mL) was added 5-bromo-2-(chloromethyl)-1,3-dimethoxybenzene (800 mg, 3.01 mmol) and K2CO3 (1.25 g, 9.04 mmol) at room temperature. After stirring overnight, the mixture was diluted with water (10 mL). The resulting solid was collected by filtration and then dried under vacuum to give the title compound. MS m / z (M+H) + : Calculated value 491.1, measured value 491.2. 1 H NMR(400 MHz,CD3OD)δ 8.53(s,1H),6.93(s,2H),5.64(d,J=1.6 Hz,2H),4.50-4.63(m,1H),3.88(s,6H),3.68-3.76(m,2H),1.61-1.86(m,2H),1.38-1.54(m,2H),0.97(t,J=7.2 Hz,3H).

[0542] Step 4: (S)-5-Azido-2-(4-bromo-2,6-dimethoxybenzyl)-N-(1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)-2H-pyrazolo[4,3-d]pyrimidin-7-amine To a solution of (S)-2-((5-azido-2-(4-bromo-2,6-dimethoxybenzyl)-2H-pyrazolo[4,3-d]pyrimidin-7-yl)amino)pentan-1-ol (900 mg, 1.832 mmol) in DMF (15 mL) was added imidazole (374 mg, 5.50 mmol) and TBDPSCl (0.565 mL, 2.198 mmol) at room temperature. After stirring overnight, the mixture was diluted with water (15 mL) and then extracted with EtOAc (15 mL x 2). The combined organic layers were washed with brine (20 mL), then dried (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-10% MeOH / DCM) to give the title compound. MS m / z (M+H) + : Calculated value 731.2, measured value 731.2.

[0543] Step 5: Benzyl (S)-4-(4-((5-azido-7-((1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate To a solution of (S)-5-azido-2-(4-bromo-2,6-dimethoxybenzyl)-N-(1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)-2H-pyrazolo[4,3-d]pyrimidin-7-amine (500 mg, 0.685 mmol) in toluene (10 mL) was added benzylpiperazine-1-carboxylate (226 mg, 1.028 mmol), RuPhos Pd G4 (58.3 mg, 0.069 mmol), and Cs2CO3 (670 mg, 2.055 mmol). The mixture was purged with N2 and then heated to 90 °C. After heating overnight, the mixture was cooled to room temperature, diluted with water (10 mL), and extracted with DCM (15 mL × 2). The combined organic layers were dried (Na2SO4) and then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-5% MeOH / DCM) to give the title compound. MS m / z (M+H) + : Calculated value 869.4, measured value 869.4.

[0544] Step 6: (S)-5-Azido-N-(1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)-2-(2,6-dimethoxy-4-(piperazin-1-yl)benzyl)-2H-pyrazolo[4,3-d]pyrimidin-7-amine To a solution of benzyl (S)-4-(4-((5-amino-7-((1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate (370 mg, 0.439 mmol) in TFE (8 mL) was added Pd / C (47 mg). The mixture was degassed (3×vacuum / N2) and then placed under an atmosphere of H2 (balloon). After 2 h, the mixture was filtered and the filtrate was concentrated to give the title compound, which was used in the next step without purification. MS m / z (M+H) + : Calculated value 735.4, measured value 735.4.

[0545] [Example 3] Preparation of Compound A-1 [ka] TIFF2026505682000104.tif170170N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (Compound A-1)

[0546] Step 1: tert-butyl 4-(4-((7-(butylamino)-5-((methoxycarbonyl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate To a stirred mixture of tert-butyl 4-(4-(hydroxymethyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate (Int. 3-1, 13.87 g, 39.4 mmol) in benzene (80 mL) under an argon atmosphere was added methyl (7-(butylamino)-1H-pyrazolo[4,3-d]pyrimidin-5-yl)carbamate (Int. 1-1, 8 g, 30.3 mmol) and cyanomethylenetributylphosphorane (10.96 g, 45.4 mmol), and the mixture was then heated to 80° C. After 3 hours, the mixture was cooled to ambient temperature, diluted with water (10 mL), and extracted with EtOAc (3×20 mL). The combined organic extracts were washed with brine (3×10 mL), dried (NaSO), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-100% EtOAc / DCM) to give the title compound. MS m / z (M+H) + : Calculated value 599.3, actual value 599.3.

[0547] Step 2: tert-butyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate To a stirred mixture of tert-butyl 4-(4-((7-(butylamino)-5-((methoxycarbonyl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate (2.6 g, 4.34 mmol) in 1,4-dioxane (5 mL) was added NaOH (10 M, 20 mL, 200 mmol) and the mixture was heated to 60° C. After 12 h, the mixture was cooled to ambient temperature, diluted with water (20 mL), and extracted with EtOAc (3×80 mL). The combined organic extracts were washed with brine (50 mL), dried (NaSO), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-10% MeOH / DCM) to give the title compound. MS m / z (M+H) + : Calculated value 541.3, measured value 541.4.

[0548] Process 3:N 7 -butyl-2-(2,6-dimethoxy-4-(piperazin-1-yl)benzyl)-2H-pyrazolo[4,3-d]pyrimidine-5,7-diamine To a stirred mixture of tert-butyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate (1.8 g, 3.33 mmol) in DCM (25 mL) was added a solution of phenol (9.40 g, 100 mmol) in DCM (5 mL) at ambient temperature. The mixture was cooled to 5° C., and then a solution of chlorotrimethylsilane (0.24 ml, 1.878 mmol) in DCM (3 mL) was added dropwise. After the addition was complete, the mixture was allowed to warm to ambient temperature. After 25 minutes, the mixture was poured into ice-cold 2N NaOH (30 mL) and then extracted with ethyl acetate (5×150 mL). The combined organic extracts were dried (NaSO) and then filtered, and the filtrate was concentrated. The crude product was subjected to reverse phase chromatography (0-60% MeCN / water with 0.5% NH4OH modifier) ​​to g...

Claims

1. 1. An immunogenic composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) A compound having the structure according to Formula I: 【Chemistry 1】 (In the formula, R ａ is H, -OH, (C １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, —O(C １ -C ６ ) selected from alkynyl, chlorine, fluorine, and —NR′R″, １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, and —O(C １ -C ６ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 4 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine; R ａ’ is H, -OH, (C １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, —O(C １ -C ６ ) selected from alkynyl, chlorine, fluorine, and —NR′R″, １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, and —O(C １ -C ６ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 4 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine; R ａ’’ is H, -OH, (C １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, —O(C １ -C ６ ) selected from alkynyl, chlorine, fluorine, and —NR′R″, １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, and —O(C １ -C ６ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 4 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine; R' and R'' are H, (C １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, and (C １ -C ６ ) alkynyl, wherein (C １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, and (C １ -C ６ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 4 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine, or R′ and R″, together with the nitrogen to which they are attached, are bonded together to form (C ３ -C ６ ) heterocycloalkyl, ３ -C ６ ) Heterocycloalkyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 4 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine; R ｂ Each occurrence of １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, —O(C １ -C ６ ) independently selected from alkynyl, chlorine, fluorine, or NR′R″; １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, and —O(C １ -C ６ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 4 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine; A is (C １ -C ６ ) alkyl, heterocycloalkyl, heterocycloalkyl-C(O)-R ｚ -, (C １ -C ４ ) alkyl-N(R ｚ )-R ｚ a carbon or nitrogen bond spacer selected from -, aryl, and heteroaryl, １ -C ６ ) alkyl, heterocycloalkyl, aryl, and heteroaryl are —OH, (C １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, —O(C １ -C ６ ) optionally substituted with 1 to 6 substituents independently selected from the group consisting of alkynyl, chlorine, fluorine, and NR'R''; １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, (C １ -C ６ ) alkynyl, —O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkenyl, and —O(C １ -C ６ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 6 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine; R ｚ Each occurrence of is independently H or (C １ -C ６ ) alkyl, B is, 【Chemistry 2】 is a functional group selected from D is (C ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl, ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 6 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine, or D is 【Transformation 3】 and Each occurrence of Z is ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl, wherein (C ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 6 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine; m is 0, 1, 2, 3, 4, or 5; n is 0, 1, 2, 3, 4, or 5. or a pharmaceutically acceptable salt thereof.

2. The compound has the structure according to formula Ia: 【Chemistry 4】 (In the formula, R' and R'' are H, (C １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, and (C １ -C ６ ) alkynyl, wherein (C １ -C ６ ) alkyl, (C １ -C ６ ) alkenyl, and (C １ -C ６ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 4 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine, or R′ and R″, together with the nitrogen to which they are attached, are bonded together to form (C ３ -C ６ ) heterocycloalkyl, ３ -C ６ ) Heterocycloalkyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 4 substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine; R ｂ Each occurrence of is -O(C １ -C ４ ) alkyl, and said —O(C １ -C ４ ) alkyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with one or two substituents independently selected from the group consisting of alkynyl, chlorine, and fluorine; A is, 【Transformation 5】 is selected from R ｚ Each occurrence of is independently H or (C １ -C ６ ) alkyl, R ｄ Each occurrence of is -OH, (C １ -C ４ ) alkyl, —O(C １ -C ４ ) independently selected from alkyl, chlorine, and fluorine; B is, 【Transformation 6】 and D is 【Transformation 7】 and a lipid chain selected from Any carbon on the lipid chain can be replaced with -OH, -O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with alkynyl, chlorine, or fluorine; 【Transformation 8】 is cis or trans stereochemistry, X １ is -O-, -C(R) ２ - or -NR-; Each occurrence of R is H, (C １ -C ４ ) alkyl, (C １ -C ４ ) alkenyl, (C １ -C ４ ) alkynyl, —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) independently selected from alkynyl, chlorine, and fluorine; m is 0, 1, or 2; n is 0, 1, 2, or 3; p is 0, 1, or 2; q is 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; s is 1, 2, 3, 4, 5, 6, 7, or 8; t is 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 2. The immunogenic composition of claim 1, comprising:

3. 3. The immunogenic composition of claim 1 or 2, wherein the composition comprises more than 20 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a specific Streptococcus pneumoniae serotype.

4. 3. The immunogenic composition of claim 1 or 2, wherein the composition comprises more than 25 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype.

5. 3. The immunogenic composition of claim 1 or 2, wherein the composition comprises 26 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae serotypes consisting of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

6. The compound has the structure according to Formula II: 【Chemistry 9】 (In the formula, R １ is (C １ -C ６ ) alkyl, and the (C １ -C ６ ) alkyl is —OH and —O(CH ３ and optionally substituted with 1 to 4 substituents independently selected from R ２ is H, methyl or —O(CH ３ ) and R ３ Each occurrence of １ -C ４ ) alkyl, (C １ -C ４ ) alkenyl, (C １ -C ４ ) alkynyl, or —O(C １ -C ４ ) alkyl, and the (C １ -C ４ ) alkyl, (C １ -C ４ ) alkenyl, (C １ -C ４ ) alkynyl, or —O(C １ -C ４ ) alkyl is —OH and —O(CH ３ and optionally substituted with one or two substituents independently selected from R ４ Each occurrence of １ -C ４ ) alkyl, (C １ -C ４ ) alkenyl, (C １ -C ４ ) alkynyl, or —O(C １ -C ４ ) alkyl, and the (C １ -C ４ ) alkyl, (C １ -C ４ ) alkenyl, (C １ -C ４ ) alkynyl, or —O(C １ -C ４ ) alkyl is —OH and —O(CH ３ and optionally substituted with one or two substituents independently selected from R ５ teeth, 【Chemistry 10】 and R ６ is (C ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl, wherein (C ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 6 substituents independently selected from alkynyl, chlorine, and fluorine; each occurrence of n is 4) 2. The immunogenic composition of claim 1, comprising:

7. The compound has the structure according to formula IIa: 【Chemistry 11】 (In the formula, R １ is butyl, said butyl being optionally substituted with one or two —OH groups; R ３ Each occurrence of is independently H or —O(CH ３ ) and R ５ teeth, 【Chemistry 12】 and R ６ is (C １０ -C ２０ ) alkyl, (C １０ -C ２０ ) alkenyl, and (C １０ -C ２０ ) alkynyl) 7. The immunogenic composition of claim 6, comprising:

8. 8. The immunogenic composition of claim 6 or 7, wherein the composition comprises more than 20 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype.

9. 8. The immunogenic composition of claim 6 or 7, wherein the composition comprises more than 25 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype.

10. 8. The immunogenic composition of claim 6 or 7, wherein the composition comprises 26 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae serotypes consisting of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

11. The compound has the structure according to Formula III: 【Chemistry 13】 (In the formula, R １ is (C １ -C ６ ) alkyl, and the (C １ -C ６ ) alkyl is —OH and —O(CH ３ and optionally substituted with 1 to 4 substituents independently selected from R ２ is H, methyl or —O(CH ３ ) and R ３ Each occurrence of １ -C ４ ) alkyl, (C １ -C ４ ) alkenyl, (C １ -C ４ ) alkynyl, or —O(C １ -C ４ ) alkyl, and the (C １ -C ４ ) alkyl, (C １ -C ４ ) alkenyl, (C １ -C ４ ) alkynyl, or —O(C １ -C ４ ) alkyl is —OH and —O(CH ３ and optionally substituted with one or two substituents independently selected from R ４ is (C ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl, wherein (C ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 6 substituents independently selected from alkynyl, chlorine, and fluorine; n is 4) 2. The immunogenic composition of claim 1, comprising:

12. The compound has the structure according to formula IIIa: 【Chemistry 14】 (In the formula, R １ is butyl, said butyl being optionally substituted with one or two —OH groups; R ３ Each occurrence of is independently H or —O(CH ３ ) and R ４ is (C １０ -C ２０ ) alkyl, (C １０ -C ２０ ) alkenyl, and (C １０ -C ２０ ) alkynyl) or a pharmaceutically acceptable salt thereof.

13. 13. The immunogenic composition of claim 11 or 12, wherein the composition comprises more than 20 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype.

14. 13. The immunogenic composition of claim 11 or 12, wherein the composition comprises more than 25 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype.

15. 13. The immunogenic composition of claim 11 or 12, wherein the composition comprises 26 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae serotypes consisting of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

16. The compound has the structure according to formula IV: 【Chemistry 15】 (In the formula, R １ is (C １ -C ６ ) alkyl, and the (C １ -C ６ ) alkyl is —OH and —O(CH ３ and optionally substituted with 1 to 4 substituents independently selected from R ２ is H, methyl or —O(CH ３ ) and R ３ Each occurrence of １ -C ４ ) alkyl, (C １ -C ４ ) alkenyl, (C １ -C ４ ) alkynyl, or —O(C １ -C ４ ) alkyl, and the (C １ -C ４ ) alkyl, (C １ -C ４ ) alkenyl, (C １ -C ４ ) alkynyl, and —O(C １ -C ４ ) alkyl is —OH and —O(CH ３ and optionally substituted with one or two substituents independently selected from R ４ Each occurrence of (C ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl, wherein (C ６ -C ２０ ) alkyl, (C ６ -C ２０ ) alkenyl, and (C ６ -C ２０ ) alkynyl is —OH, —O(C １ -C ４ ) alkyl, —O(C １ -C ４ ) alkenyl, —O(C １ -C ４ ) optionally substituted with 1 to 6 substituents independently selected from alkynyl, chlorine, or fluorine; n is 4) 2. The immunogenic composition of claim 1, comprising:

17. The compound has the structure according to formula IVa: 【Chemistry 16】 (In the formula, R １ is butyl, said butyl being optionally substituted with one or two —OH groups; R ３ Each occurrence of is independently H or —O(CH ３ ) and R ４ Each occurrence of (C １０ -C ２０ ) alkyl, (C １０ -C ２０ ) alkenyl, and (C １０ -C ２０ ) alkynyl) 17. The immunogenic composition of claim 16, comprising:

18. 18. The immunogenic composition of claim 16 or 17, wherein the composition comprises more than 20 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype.

19. 18. The immunogenic composition of claim 16 or 17, wherein the composition comprises more than 25 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype.

20. 18. The immunogenic composition of claim 16 or 17, wherein the composition comprises 26 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae serotypes consisting of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

21. The compound is (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, (S)—N-(5-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)tetradecanamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)oleamide, (9Z,12Z)-N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)octadeca-9,12-dienamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepan-1-yl)-5-oxopentyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperidin-1-yl)-5-oxopentyl)stearamide, N-(5-(3-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)azetidin-1-yl)-5-oxopentyl)stearamide, 1-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperidine-4-carboxamide, (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-hexadecylcyclobutane-1-carboxamide, N-(3-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-3-oxopropyl)stearamide, N-(7-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-7-oxoheptyl)stearamide, N-(3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)cyclobutyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-4,4-dimethyl-5-oxopentyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-3-methyl-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, (9Z,12Z)-N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)octadeca-9,12-dienamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)tetradecanamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)oleamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)-4-oxobutyl)stearamide, (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)carbamate, 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperazine-1-carboxamide, and 3-stearamidopropyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazine-1-carboxylate, The immunogenic composition of any one of claims 1 to 20, wherein the immunogenic composition is selected from the group consisting of:

22. The compound is N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, The immunogenic composition of any one of claims 1 to 21, which is a pharmaceutically acceptable salt thereof.

23. The immunogenic composition of any one of claims 1 to 22, further comprising: (iii) sorbitan trioleate (SPAN-85); (iv) polysorbate-20 (PS-20) or polysorbate-80 (PS-80); and (v) squalene.

24. 24. The immunogenic composition of claim 23, wherein the compound is N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidin-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

25. The immunogenic composition of claim 24, wherein the composition comprises PS-20.

26. 26. The immunogenic composition of any one of claims 23 to 25, wherein the composition comprises more than 20 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype.

27. 26. The immunogenic composition of any one of claims 23 to 25, wherein the composition comprises more than 25 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype.

28. 26. The immunogenic composition of any one of claims 23 to 25, wherein the composition comprises 26 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a polysaccharide of a particular Streptococcus pneumoniae serotype conjugated to a carrier protein, the Streptococcus pneumoniae serotypes consisting of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

29. (vi) The immunogenic composition of any one of claims 1 to 28, further comprising a pharmaceutically acceptable carrier.

30. The immunogenic composition of any one of claims 1 to 29, wherein the carrier protein is CRM197.

31. 31. The immunogenic composition of any one of claims 23 to 30, wherein the concentration of the compound is 0.1 μg / mL to 100 μg / mL, the concentration of SPAN-85 is 0.01 mg / mL to 50 mg / mL, the concentration of PS-20 or PS-80 is 0.01 mg / mL to 50 mg / mL, and the concentration of squalene is 0.02 mg / mL to 20 mg / mL.

32. 32. A method of treating or preventing pneumococcal disease in a human patient, comprising administering to said patient an immunogenic composition according to any one of claims 1 to 31.

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